LOCUS       Exported               11106 bp ds-DNA     circular SYN 13-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 RDS1a sequence 
            ATTCAATACGAAATGTGTGC in yeast chromosome 3.
ACCESSION   .
VERSION     .
KEYWORDS    p426_Cas9_gRNA-RDS1a
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 13, 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     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    424..445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        460..490
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    498..514
                     /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     503..525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     522..538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     522..538
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     557..577
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        559..577
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        804..879
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        965..1669
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1679..5782
                     /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             5795..5815
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5831..6078
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6096..6115)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6097..6115)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6125..6142)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6125..6141)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6134..6156)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6282..6737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6369..6388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6579..6600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6868..7671)
                     /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(7672..7887)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7911..7930)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8030..8052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8090..8108)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8149..9491
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8481..8528
                     /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        9518..9622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9623..10483
                     /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(9841..9860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10654..136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      541 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctagtata
      601 ctctttcttc aacaattaaa tactctcggt agccaagttg gtttaaggcg caagactgta
      661 atttatcact acgaaatctt gagatcgggc gttcgactcg cccccgggag agatggccgg
      721 catggtccca gcctcctcgc tggcgccggc tgggcaacac cttcgggtgg cgaatgggac
      781 tttattcaat acgaaatgtg tgcgttttag agctagaaat agcaagttaa aataaggcta
      841 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttattt tttgtcacta
      901 ttgttatgta aaatgccacc tctgacagta tggaacgcaa acttctgtct agtggatagt
      961 cgacatcctt ttgttgtttc cgggtgtaca atatggactt cctcttttct ggcaaccaaa
     1021 cccatacatc gggattccta taataccttc gttggtctcc ctaacatgta ggtggcggag
     1081 gggagatata caatagaaca gataccagac aagacataat gggctaaaca agactacacc
     1141 aattacactg cctcattgat ggtggtacat aacgaactaa tactgtagcc ctagacttga
     1201 tagccatcat catatcgaag tttcactacc ctttttccat ttgccatcta ttgaagtaat
     1261 aataggcgca tgcaacttct tttctttttt tttcttttct ctctcccccg ttgttgtctc
     1321 accatatccg caatgacaaa aaaatgatgg aagacactaa aggaaaaaat taacgacaaa
     1381 gacagcacca acagatgtcg ttgttccaga gctgatgagg ggtatctcga agcacacgaa
     1441 actttttcct tccttcattc acgcacacta ctctctaatg agcaacggta tacggccttc
     1501 cttccagtta cttgaatttg aaataaaaaa aagtttgctg tcttgctatc aagtataaat
     1561 agacctgcaa ttattaatct tttgtttcct cgtcattgtt ctcgttccct ttcttccttg
     1621 tttctttttc tgcacaatat ttcaagctat accaagcata caatcaactt ggaaaaaaat
     1681 ggacaagaag tactccattg ggctcgatat cggcacaaac agcgtcggtt gggccgtcat
     1741 tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca
     1801 cagcataaag aagaacctca ttggcgccct cctgttcgac tccggggaga cggccgaagc
     1861 cacgcggctc aaaagaacag cacggcgcag atatacccgc agaaagaatc ggatctgcta
     1921 cctgcaggag atctttagta atgagatggc taaggtggat gactctttct tccataggct
     1981 ggaggagtcc tttttggtgg aggaggataa aaagaacgag cgccacccaa tctttggcaa
     2041 tatcgtggac gaggtggcgt accatgaaaa gtacccaacc atatatcatc tgaggaagaa
     2101 gcttgtagac agtactgata aggctgactt gcggttgatc tatctcgcgc tggcgcatat
     2161 gatcaaattt cggggacact tcctcatcga gggggacctg aacccagaca acagcgatgt
     2221 cgacaaactc tttatccaac tggttcagac ttacaatcag cttttcgaag agaacccgat
     2281 caacgcatcc ggagttgacg ccaaagcaat cctgagcgct aggctgtcca aatcccggcg
     2341 gctcgaaaac ctcatcgcac agctccctgg ggagaagaag aacggcctgt ttggtaatct
     2401 tatcgccctg tcactcgggc tgacccccaa ctttaaatct aacttcgacc tggccgaaga
     2461 tgccaagctt caactgagca aagacaccta cgatgatgat ctcgacaatc tgctggccca
     2521 gatcggcgac cagtacgcag accttttttt ggcggcaaag aacctgtcag acgccattct
     2581 gctgagtgat attctgcgag tgaacacgga gatcaccaaa gctccgctga gcgctagtat
     2641 gatcaagcgc tatgatgagc accaccaaga cttgactttg ctgaaggccc ttgtcagaca
     2701 gcaactgcct gagaagtaca aggaaatttt cttcgatcag tctaaaaatg gctacgccgg
     2761 atacattgac ggcggagcaa gccaggagga attttacaaa tttattaagc ccatcttgga
     2821 aaaaatggac ggcaccgagg agctgctggt aaagcttaac agagaagatc tgttgcgcaa
     2881 acagcgcact ttcgacaatg gaagcatccc ccaccagatt cacctgggcg aactgcacgc
     2941 tatcctcagg cggcaagagg atttctaccc ctttttgaaa gataacaggg aaaagattga
     3001 gaaaatcctc acatttcgga taccctacta tgtaggcccc ctcgcccggg gaaattccag
     3061 attcgcgtgg atgactcgca aatcagaaga gaccatcact ccctggaact tcgaggaagt
     3121 cgtggataag ggggcctctg cccagtcctt catcgaaagg atgactaact ttgataaaaa
     3181 tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg tacgagtact tcacagttta
     3241 taacgagctc accaaggtca aatacgtcac agaagggatg agaaagccag cattcctgtc
     3301 tggagagcag aagaaagcta tcgtggacct cctcttcaag acgaaccgga aagttaccgt
     3361 gaaacagctc aaagaagact atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag
     3421 cggagtggag gatcgcttca acgcatccct gggaacgtat cacgatctcc tgaaaatcat
     3481 taaagacaag gacttcctgg acaatgagga gaacgaggac attcttgagg acattgtcct
     3541 cacccttacg ttgtttgaag atagggagat gattgaagaa cgcttgaaaa cttacgctca
     3601 tctcttcgac gacaaagtca tgaaacagct caagaggcgc cgatatacag gatgggggcg
     3661 gctgtcaaga aaactgatca atgggatccg agacaagcag agtggaaaga caatcctgga
     3721 ttttcttaag tccgatggat ttgccaaccg gaacttcatg cagttgatcc atgatgactc
     3781 tctcaccttt aaggaggaca tccagaaagc acaagtttct ggccaggggg acagtcttca
     3841 cgagcacatc gctaatcttg caggtagccc agctatcaaa aagggaatac tgcagaccgt
     3901 taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat aagcccgaga atatcgttat
     3961 cgagatggcc cgagagaacc aaactaccca gaagggacag aagaacagta gggaaaggat
     4021 gaagaggatt gaagagggta taaaagaact ggggtcccaa atccttaagg aacacccagt
     4081 tgaaaacacc cagcttcaga atgagaagct ctacctgtac tacctgcaga acggcaggga
     4141 catgtacgtg gatcaggaac tggacatcaa tcggctctcc gactacgacg tggatcatat
     4201 cgtgccccag tcttttctca aagatgattc tattgataat aaagtgttga caagatccga
     4261 taaaaataga gggaagagtg ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa
     4321 ttattggcgg cagctgctga acgccaaact gatcacacaa cggaagttcg ataatctgac
     4381 taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc ggcttcatca aaaggcagct
     4441 tgttgagaca cgccagatca ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac
     4501 caagtacgat gaaaatgaca aactgattcg agaggtgaaa gttattactc tgaagtctaa
     4561 gctggtctca gatttcagaa aggactttca gttttataag gtgagagaga tcaacaatta
     4621 ccaccatgcg catgatgcct acctgaatgc agtggtaggc actgcactta tcaaaaaata
     4681 tcccaagctt gaatctgaat ttgtttacgg agactataaa gtgtacgatg ttaggaaaat
     4741 gatcgcaaag tctgagcagg aaataggcaa ggccaccgct aagtacttct tttacagcaa
     4801 tattatgaat tttttcaaga ccgagattac actggccaat ggagagattc ggaagcgacc
     4861 acttatcgaa acaaacggag aaacaggaga aatcgtgtgg gacaagggta gggatttcgc
     4921 gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca
     4981 gaccggaggc ttctccaagg aaagtatcct cccgaaaagg aacagcgaca agctgatcgc
     5041 acgcaaaaaa gattgggacc ccaagaaata cggcggattc gattctccta cagtcgctta
     5101 cagtgtactg gttgtggcca aagtggagaa agggaagtct aaaaaactca aaagcgtcaa
     5161 ggaactgctg ggcatcacaa tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt
     5221 tctcgaggcg aaaggatata aagaggtcaa aaaagacctc atcattaagc ttcccaagta
     5281 ctctctcttt gagcttgaaa acggccggaa acgaatgctc gctagtgcgg gcgagctgca
     5341 gaaaggtaac gagctggcac tgccctctaa atacgttaat ttcttgtatc tggccagcca
     5401 ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag aagcagctgt tcgtggaaca
     5461 acacaaacac taccttgatg agatcatcga gcaaataagc gaattctcca aaagagtgat
     5521 cctcgccgac gctaacctcg ataaggtgct ttctgcttac aataagcaca gggataagcc
     5581 catcagggag caggcagaaa acattatcta cttgtttact ctgaccaact tgggcgcgcc
     5641 tgcagccttc aagtacttcg acaccaccat agacagaaag cggtacacct ctacaaagga
     5701 ggtcctggac gccacactga ttcatcagtc aattacgggg ctctatgaaa caagaatcga
     5761 cctctctcag ctcggtggag acagcagggc tgaccccaag aagaagagga aggtgtgatc
     5821 tcttctcgag tcatgtaatt agttatgtca cgcttacgtt cacgccctcc ccccacatcc
     5881 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     5941 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     6001 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     6061 tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     6121 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     6181 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     6241 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     6301 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6361 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6421 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6481 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6541 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6601 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6661 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6721 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6781 caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     6841 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     6901 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     6961 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     7021 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     7081 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     7141 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     7201 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     7261 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     7321 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     7381 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     7441 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     7501 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     7561 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     7621 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     7681 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     7741 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     7801 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     7861 agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc
     7921 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     7981 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     8041 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     8101 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagtatga tccaatatca
     8161 aaggaaatga tagcattgaa ggatgagact aatccaattg aggagtggca gcatatagaa
     8221 cagctaaagg gtagtgctga aggaagcata cgataccccg catggaatgg gataatatca
     8281 caggaggtac tagactacct ttcatcctac ataaatagac gcatataagt acgcatttaa
     8341 gcataaacac gcactatgcc gttcttctca tgtatatata tatacaggca acacgcagat
     8401 ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg
     8461 ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt cctattctct agaaagtata
     8521 ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa gacgcacttt caaaaaacca
     8581 aaaacgcacc ggactgtaac gagctactaa aatattgcga ataccgcttc cacaaacatt
     8641 gctcaaaagt atctctttgc tatatatctc tgtgctatat ccctatataa cctacccatc
     8701 cacctttcgc tccttgaact tgcatctaaa ctcgacctct acatttttta tgtttatctc
     8761 tagtattact ctttagacaa aaaaattgta gtaagaacta ttcatagagt gaatcgaaaa
     8821 caatacgaaa atgtaaacat ttcctatacg tagtatatag agacaaaata gaagaaaccg
     8881 ttcataattt tctgaccaat gaagaatcat caacgctatc actttctgtt cacaaagtat
     8941 gcgcaatcca catcggtata gaatataatc ggggatgcct ttatcttgaa aaaatgcacc
     9001 cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt caggcttttt ttatggaaga
     9061 gaaaatagac accaaagtag ccttcttcta accttaacgg acctacagtg caaaaagtta
     9121 tcaagagact gcattataga gcgcacaaag gagaaaaaaa gtaatctaag atgctttgtt
     9181 agaaaaatag cgctctcggg atgcattttt gtagaacaaa aaagaagtat agattctttg
     9241 ttggtaaaat agcgctctcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     9301 tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt acaaaaatga agcacagatt
     9361 cttcgttggt aaaatagcgc tttcgcgttg catttctgtt ctgtaaaaat gcagctcaga
     9421 ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttctacaaa atgaagcaca
     9481 gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     9541 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     9601 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     9661 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     9721 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     9781 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     9841 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     9901 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     9961 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
    10021 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
    10081 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
    10141 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
    10201 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
    10261 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
    10321 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
    10381 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
    10441 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
    10501 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
    10561 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
    10621 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
    10681 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
    10741 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
    10801 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
    10861 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
    10921 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
    10981 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
    11041 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
    11101 cggcag
//
