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 ARS1114a sequence 
            CTTGTGAAACAAATAATTGG in yeast chromosome 11..
ACCESSION   .
VERSION     .
KEYWORDS    p426_Cas9_gRNA-ARS1114a
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"
     promoter        complement(23..238)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(262..281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     381..403
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(441..459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      500..1842
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    832..879
                     /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        1869..1973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1974..2834
                     /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(2192..2211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3005..3593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3494..3513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3747..3764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3881..3902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3917..3947
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3955..3971
                     /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     3960..3982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3979..3995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3979..3995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4014..4034
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4016..4034
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4261..4336
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4422..5126
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5136..9239
                     /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             9252..9272
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9288..9535
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9553..9572)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9554..9572)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9582..9599)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9582..9598)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9591..9613)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9739..10194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9826..9845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10036..10057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(10325..22)
                     /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"
ORIGIN
        1 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca
       61 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
      121 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
      181 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
      241 ggtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
      301 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      361 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      421 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
      481 aataatggtt tcttagtatg atccaatatc aaaggaaatg atagcattga aggatgagac
      541 taatccaatt gaggagtggc agcatataga acagctaaag ggtagtgctg aaggaagcat
      601 acgatacccc gcatggaatg ggataatatc acaggaggta ctagactacc tttcatccta
      661 cataaataga cgcatataag tacgcattta agcataaaca cgcactatgc cgttcttctc
      721 atgtatatat atatacaggc aacacgcaga tataggtgcg acgtgaacag tgagctgtat
      781 gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt tgaagttcct
      841 attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt gaaaaccaaa
      901 agcgctctga agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta
      961 aaatattgcg aataccgctt ccacaaacat tgctcaaaag tatctctttg ctatatatct
     1021 ctgtgctata tccctatata acctacccat ccacctttcg ctccttgaac ttgcatctaa
     1081 actcgacctc tacatttttt atgtttatct ctagtattac tctttagaca aaaaaattgt
     1141 agtaagaact attcatagag tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac
     1201 gtagtatata gagacaaaat agaagaaacc gttcataatt ttctgaccaa tgaagaatca
     1261 tcaacgctat cactttctgt tcacaaagta tgcgcaatcc acatcggtat agaatataat
     1321 cggggatgcc tttatcttga aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg
     1381 ggaagtggag tcaggctttt tttatggaag agaaaataga caccaaagta gccttcttct
     1441 aaccttaacg gacctacagt gcaaaaagtt atcaagagac tgcattatag agcgcacaaa
     1501 ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata gcgctctcgg gatgcatttt
     1561 tgtagaacaa aaaagaagta tagattcttt gttggtaaaa tagcgctctc gcgttgcatt
     1621 tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc
     1681 atttttgttt tacaaaaatg aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt
     1741 gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc
     1801 gttgcatttt tgttctacaa aatgaagcac agatgcttcg ttcaggtggc acttttcggg
     1861 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1921 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2161 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2281 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2521 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2581 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2821 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     3841 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     3901 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     3961 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     4021 accctcacta aagggaacaa aagctggagc tagtatactc tttcttcaac aattaaatac
     4081 tctcggtagc caagttggtt taaggcgcaa gactgtaatt tatcactacg aaatcttgag
     4141 atcgggcgtt cgactcgccc ccgggagaga tggccggcat ggtcccagcc tcctcgctgg
     4201 cgccggctgg gcaacacctt cgggtggcga atgggacttt cttgtgaaac aaataattgg
     4261 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     4321 ggcaccgagt cggtgctttt tttatttttt gtcactattg ttatgtaaaa tgccacctct
     4381 gacagtatgg aacgcaaact tctgtctagt ggatagtcga catccttttg ttgtttccgg
     4441 gtgtacaata tggacttcct cttttctggc aaccaaaccc atacatcggg attcctataa
     4501 taccttcgtt ggtctcccta acatgtaggt ggcggagggg agatatacaa tagaacagat
     4561 accagacaag acataatggg ctaaacaaga ctacaccaat tacactgcct cattgatggt
     4621 ggtacataac gaactaatac tgtagcccta gacttgatag ccatcatcat atcgaagttt
     4681 cactaccctt tttccatttg ccatctattg aagtaataat aggcgcatgc aacttctttt
     4741 cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa
     4801 atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg
     4861 ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg
     4921 cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa
     4981 taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt
     5041 gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc
     5101 aagctatacc aagcatacaa tcaacttgga aaaaaatgga caagaagtac tccattgggc
     5161 tcgatatcgg cacaaacagc gtcggttggg ccgtcattac ggacgagtac aaggtgccga
     5221 gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg
     5281 gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac
     5341 ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg
     5401 agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg
     5461 aggataaaaa gaacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc
     5521 atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg
     5581 ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc
     5641 tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg
     5701 ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca
     5761 aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc
     5821 tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga
     5881 cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag
     5941 acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc
     6001 tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga
     6061 acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc
     6121 accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg
     6181 aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc
     6241 aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc
     6301 tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa
     6361 gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt
     6421 tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac
     6481 cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat
     6541 cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc
     6601 agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc
     6661 ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat
     6721 acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg
     6781 tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt
     6841 tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg
     6901 catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca
     6961 atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata
     7021 gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga
     7081 aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg
     7141 ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg
     7201 ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc
     7261 agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag
     7321 gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca
     7381 aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa
     7441 ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa
     7501 aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg
     7561 agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg
     7621 acatcaatcg gctctccgac tacgacgtgg atcatatcgt gccccagtct tttctcaaag
     7681 atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg aagagtgata
     7741 acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg
     7801 ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt
     7861 ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca
     7921 agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac
     7981 tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg
     8041 actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc
     8101 tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg
     8161 tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa
     8221 taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg
     8281 agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa
     8341 caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca
     8401 tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa
     8461 gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca
     8521 agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag
     8581 tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca
     8641 tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag
     8701 aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg
     8761 gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc
     8821 cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc
     8881 ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga
     8941 tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata
     9001 aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca
     9061 ttatctactt gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca
     9121 ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc
     9181 atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagaca
     9241 gcagggctga ccccaagaag aagaggaagg tgtgatctct tctcgagtca tgtaattagt
     9301 tatgtcacgc ttacgttcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt
     9361 tagacaacct gaagtctagg tccctattta tttttttata gttatgttag tattaagaac
     9421 gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat
     9481 tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg
     9541 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     9601 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     9661 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     9721 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     9781 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     9841 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     9901 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     9961 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
    10021 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
    10081 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
    10141 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
    10201 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
    10261 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
    10321 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
    10381 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
    10441 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
    10501 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
    10561 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
    10621 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
    10681 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
    10741 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
    10801 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
    10861 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
    10921 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
    10981 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
    11041 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
    11101 cacgtt
//
