LOCUS       sequence-193446-       11106 bp    DNA     circular SYN 15-DEC-2025
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 ARS1622b sequence 
            GTCACGTTCCTGAGGTTACT in yeast chromosome 16..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/nar/gkw1023"; journalName: "Nucleic Acids Res"; date: 
            "2017-01-9- 9"; volume: "45"; issue: "1"; pages: "496-508"
COMMENT     Sequence Label: p426_Cas9_gRNA-ARS1622b
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     165..187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(225..243)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      284..1626
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    616..663
                     /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        1653..1757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1758..1826,1827..2618)
                     /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(1976..1995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2789..3377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3278..3297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3531..3548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3665..3686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3701..3706,3707..3724,3725..3731)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3755
                     /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     3744..3766
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3763..3779
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3763..3779
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3798..3818
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3800..3818
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4045..4120
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4206..4910
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4920..9023
                     /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             9036..9056
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9072..9319
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9337..9356)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9338..9356)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9366..9383)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9366..9382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9375..9397)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9523..9978
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9610..9629)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9820..9841
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(10109..10912)
                     /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(join(10913..11106,1..22))
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 aaaaaaaaaa tgatgaattg aaaaggtggt atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      121 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
      181 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
      241 atttttatag gttaatgtca tgataataat ggtttcttag tatgatccaa tatcaaagga
      301 aatgatagca ttgaaggatg agactaatcc aattgaggag tggcagcata tagaacagct
      361 aaagggtagt gctgaaggaa gcatacgata ccccgcatgg aatgggataa tatcacagga
      421 ggtactagac tacctttcat cctacataaa tagacgcata taagtacgca tttaagcata
      481 aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
      541 tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
      601 tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac
      661 ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
      721 gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
      781 aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
      841 ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta
      901 ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata
      961 cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat
     1021 aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca
     1081 atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag
     1141 cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa
     1201 tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag
     1261 agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa
     1321 aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt
     1381 aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga ttctttgttt
     1441 gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac agattcttcg
     1501 ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     1561 tgtttgaaaa attagcgctc tcgcgttgca tttttgttct acaaaatgaa gcacagatgc
     1621 ttcgttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1681 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1741 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1801 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1861 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1921 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1981 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2041 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2101 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2161 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2221 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2281 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2341 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2401 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2461 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2521 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2581 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2641 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2701 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2761 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2821 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2881 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2941 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3001 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3061 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3121 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3181 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3241 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3301 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3361 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3421 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3481 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3541 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3601 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3661 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     3721 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     3781 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctagtat
     3841 actctttctt caacaattaa atactctcgg tagccaagtt ggtttaaggc gcaagactgt
     3901 aatttatcac tacgaaatct tgagatcggg cgttcgactc gcccccggga gagatggccg
     3961 gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca ccttcgggtg gcgaatggga
     4021 ctttgtcacg ttcctgaggt tactgtttta gagctagaaa tagcaagtta aaataaggct
     4081 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttatt ttttgtcact
     4141 attgttatgt aaaatgccac ctctgacagt atggaacgca aacttctgtc tagtggatag
     4201 tcgacatcct tttgttgttt ccgggtgtac aatatggact tcctcttttc tggcaaccaa
     4261 acccatacat cgggattcct ataatacctt cgttggtctc cctaacatgt aggtggcgga
     4321 ggggagatat acaatagaac agataccaga caagacataa tgggctaaac aagactacac
     4381 caattacact gcctcattga tggtggtaca taacgaacta atactgtagc cctagacttg
     4441 atagccatca tcatatcgaa gtttcactac cctttttcca tttgccatct attgaagtaa
     4501 taataggcgc atgcaacttc ttttcttttt ttttcttttc tctctccccc gttgttgtct
     4561 caccatatcc gcaatgacaa aaaaatgatg gaagacacta aaggaaaaaa ttaacgacaa
     4621 agacagcacc aacagatgtc gttgttccag agctgatgag gggtatctcg aagcacacga
     4681 aactttttcc ttccttcatt cacgcacact actctctaat gagcaacggt atacggcctt
     4741 ccttccagtt acttgaattt gaaataaaaa aaagtttgct gtcttgctat caagtataaa
     4801 tagacctgca attattaatc ttttgtttcc tcgtcattgt tctcgttccc tttcttcctt
     4861 gtttcttttt ctgcacaata tttcaagcta taccaagcat acaatcaact tggaaaaaaa
     4921 tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggt tgggccgtca
     4981 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
     5041 acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag acggccgaag
     5101 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
     5161 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
     5221 tggaggagtc ctttttggtg gaggaggata aaaagaacga gcgccaccca atctttggca
     5281 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
     5341 agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcata
     5401 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
     5461 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa gagaacccga
     5521 tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     5581 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     5641 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
     5701 atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
     5761 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
     5821 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
     5881 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
     5941 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
     6001 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
     6061 aaaaaatgga cggcaccgag gagctgctgg taaagcttaa cagagaagat ctgttgcgca
     6121 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
     6181 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
     6241 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgcccgg ggaaattcca
     6301 gattcgcgtg gatgactcgc aaatcagaag agaccatcac tccctggaac ttcgaggaag
     6361 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
     6421 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
     6481 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
     6541 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
     6601 tgaaacagct caaagaagac tatttcaaaa agattgaatg tttcgactct gttgaaatca
     6661 gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca
     6721 ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc
     6781 tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc
     6841 atctcttcga cgacaaagtc atgaaacagc tcaagaggcg ccgatataca ggatgggggc
     6901 ggctgtcaag aaaactgatc aatgggatcc gagacaagca gagtggaaag acaatcctgg
     6961 attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact
     7021 ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg gacagtcttc
     7081 acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg
     7141 ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta
     7201 tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga
     7261 tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag
     7321 ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg
     7381 acatgtacgt ggatcaggaa ctggacatca atcggctctc cgactacgac gtggatcata
     7441 tcgtgcccca gtcttttctc aaagatgatt ctattgataa taaagtgttg acaagatccg
     7501 ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa
     7561 attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga
     7621 ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc
     7681 ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca
     7741 ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta
     7801 agctggtctc agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt
     7861 accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat
     7921 atcccaagct tgaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa
     7981 tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca
     8041 atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt cggaagcgac
     8101 cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg
     8161 cgacagtccg gaaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac
     8221 agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg
     8281 cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt
     8341 acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca
     8401 aggaactgct gggcatcaca atcatggagc gatcaagctt cgaaaaaaac cccatcgact
     8461 ttctcgaggc gaaaggatat aaagaggtca aaaaagacct catcattaag cttcccaagt
     8521 actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc
     8581 agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc
     8641 actatgaaaa gctcaaaggg tctcccgaag ataatgagca gaagcagctg ttcgtggaac
     8701 aacacaaaca ctaccttgat gagatcatcg agcaaataag cgaattctcc aaaagagtga
     8761 tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc
     8821 ccatcaggga gcaggcagaa aacattatct acttgtttac tctgaccaac ttgggcgcgc
     8881 ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg
     8941 aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg
     9001 acctctctca gctcggtgga gacagcaggg ctgaccccaa gaagaagagg aaggtgtgat
     9061 ctcttctcga gtcatgtaat tagttatgtc acgcttacgt tcacgccctc cccccacatc
     9121 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     9181 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     9241 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     9301 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     9361 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     9421 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     9481 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     9541 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     9601 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     9661 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     9721 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     9781 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     9841 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     9901 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     9961 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
    10021 acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
    10081 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
    10141 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
    10201 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
    10261 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
    10321 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
    10381 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
    10441 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
    10501 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
    10561 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
    10621 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
    10681 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
    10741 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
    10801 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
    10861 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
    10921 ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
    10981 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
    11041 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
    11101 aagaat
//
