LOCUS       sequence-193444-       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 YOLCd1b sequence 
            GACTAGTTAAGCGAGCATGT in yeast chromosome 15..
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-YOLCd1b
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..71)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(225..813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(305..324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(984..1775,1776..1844))
                     /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     1607..1626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1845..1949)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1976..3318
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2939..2986)
                     /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)."
     primer_bind     3359..3377
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3415..3437)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3537..3556
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3580..3795
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3796..4599
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(4730..5185)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4867..4888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5079..5098
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5311..5333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5325..5342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5326..5342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5352..5371
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5352..5370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5389..5636)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(5652..5672)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5685..9788)
                     /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"
     promoter        complement(9798..10502)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_RNA        complement(10588..10663)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(10890..10910)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10890..10908)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10929..10945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10929..10945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10942..10964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10953..10969
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(10977..10983,10984..11001,11002..11007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11022..11043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
       61 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1261 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaacga
     1981 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2041 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     2101 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     2161 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     2221 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     2281 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     2341 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     2401 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     2461 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     2521 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     2581 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     2641 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     2701 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     2761 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     2821 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     2881 cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga
     2941 agttcctata ctttctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa
     3001 aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc
     3061 acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt
     3121 ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt agtctagtac
     3181 ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag cactaccctt
     3241 tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca atgctatcat
     3301 ttcctttgat attggatcat actaagaaac cattattatc atgacattaa cctataaaaa
     3361 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3421 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3481 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3541 atcagagcag attgtactga gagtgcacca taccaccttt tcaattcatc attttttttt
     3601 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
     3661 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     3721 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg
     3781 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     3841 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     3901 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     3961 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     4021 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
     4081 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     4141 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     4201 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     4261 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     4321 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     4381 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa
     4441 cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga
     4501 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     4561 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     4621 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attaccctat
     4681 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     4741 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4801 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4861 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4921 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4981 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5041 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5101 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5161 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     5221 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     5281 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     5341 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccggccgc aaattaaagc
     5401 cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac
     5461 gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat
     5521 aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt
     5581 tagagcggat gtggggggag ggcgtgaacg taagcgtgac ataactaatt acatgactcg
     5641 agaagagatc acaccttcct cttcttcttg gggtcagccc tgctgtctcc accgagctga
     5701 gagaggtcga ttcttgtttc atagagcccc gtaattgact gatgaatcag tgtggcgtcc
     5761 aggacctcct ttgtagaggt gtaccgcttt ctgtctatgg tggtgtcgaa gtacttgaag
     5821 gctgcaggcg cgcccaagtt ggtcagagta aacaagtaga taatgttttc tgcctgctcc
     5881 ctgatgggct tatccctgtg cttattgtaa gcagaaagca ccttatcgag gttagcgtcg
     5941 gcgaggatca ctcttttgga gaattcgctt atttgctcga tgatctcatc aaggtagtgt
     6001 ttgtgttgtt ccacgaacag ctgcttctgc tcattatctt cgggagaccc tttgagcttt
     6061 tcatagtggc tggccagata caagaaatta acgtatttag agggcagtgc cagctcgtta
     6121 cctttctgca gctcgcccgc actagcgagc attcgtttcc ggccgttttc aagctcaaag
     6181 agagagtact tgggaagctt aatgatgagg tcttttttga cctctttata tcctttcgcc
     6241 tcgagaaagt cgatggggtt tttttcgaag cttgatcgct ccatgattgt gatgcccagc
     6301 agttccttga cgcttttgag ttttttagac ttccctttct ccactttggc cacaaccagt
     6361 acactgtaag cgactgtagg agaatcgaat ccgccgtatt tcttggggtc ccaatctttt
     6421 ttgcgtgcga tcagcttgtc gctgttcctt ttcgggagga tactttcctt ggagaagcct
     6481 ccggtctgta cttcggtctt tttaacgatg ttcacctgcg gcatggacag gaccttccgg
     6541 actgtcgcga aatccctacc cttgtcccac acgatttctc ctgtttctcc gtttgtttcg
     6601 ataagtggtc gcttccgaat ctctccattg gccagtgtaa tctcggtctt gaaaaaattc
     6661 ataatattgc tgtaaaagaa gtacttagcg gtggccttgc ctatttcctg ctcagacttt
     6721 gcgatcattt tcctaacatc gtacacttta tagtctccgt aaacaaattc agattcaagc
     6781 ttgggatatt ttttgataag tgcagtgcct accactgcat tcaggtaggc atcatgcgca
     6841 tggtggtaat tgttgatctc tctcacctta taaaactgaa agtcctttct gaaatctgag
     6901 accagcttag acttcagagt aataactttc acctctcgaa tcagtttgtc attttcatcg
     6961 tacttggtgt tcatgcgtga atcgagaatt tgggccacgt gcttggtgat ctggcgtgtc
     7021 tcaacaagct gccttttgat gaagccggct ttatccaact cagacaggcc acctcgttca
     7081 gccttagtca gattatcgaa cttccgttgt gtgatcagtt tggcgttcag cagctgccgc
     7141 caataatttt tcattttctt gacaacttct tctgagggga cgttatcact cttccctcta
     7201 tttttatcgg atcttgtcaa cactttatta tcaatagaat catctttgag aaaagactgg
     7261 ggcacgatat gatccacgtc gtagtcggag agccgattga tgtccagttc ctgatccacg
     7321 tacatgtccc tgccgttctg caggtagtac aggtagagct tctcattctg aagctgggtg
     7381 ttttcaactg ggtgttcctt aaggatttgg gaccccagtt cttttatacc ctcttcaatc
     7441 ctcttcatcc tttccctact gttcttctgt cccttctggg tagtttggtt ctctcgggcc
     7501 atctcgataa cgatattctc gggcttatgc cttcccatta ctttgacgag ttcatccacg
     7561 accttaacgg tctgcagtat tccctttttg atagctgggc tacctgcaag attagcgatg
     7621 tgctcgtgaa gactgtcccc ctggccagaa acttgtgctt tctggatgtc ctccttaaag
     7681 gtgagagagt catcatggat caactgcatg aagttccggt tggcaaatcc atcggactta
     7741 agaaaatcca ggattgtctt tccactctgc ttgtctcgga tcccattgat cagttttctt
     7801 gacagccgcc cccatcctgt atatcggcgc ctcttgagct gtttcatgac tttgtcgtcg
     7861 aagagatgag cgtaagtttt caagcgttct tcaatcatct ccctatcttc aaacaacgta
     7921 agggtgagga caatgtcctc aagaatgtcc tcgttctcct cattgtccag gaagtccttg
     7981 tctttaatga ttttcaggag atcgtgatac gttcccaggg atgcgttgaa gcgatcctcc
     8041 actccgctga tttcaacaga gtcgaaacat tcaatctttt tgaaatagtc ttctttgagc
     8101 tgtttcacgg taactttccg gttcgtcttg aagaggaggt ccacgatagc tttcttctgc
     8161 tctccagaca ggaatgctgg ctttctcatc ccttctgtga cgtatttgac cttggtgagc
     8221 tcgttataaa ctgtgaagta ctcgtacagc agagagtgtt taggaagcac cttttcgtta
     8281 ggcagatttt tatcaaagtt agtcatcctt tcgatgaagg actgggcaga ggccccctta
     8341 tccacgactt cctcgaagtt ccagggagtg atggtctctt ctgatttgcg agtcatccac
     8401 gcgaatctgg aatttccccg ggcgaggggg cctacatagt agggtatccg aaatgtgagg
     8461 attttctcaa tcttttccct gttatctttc aaaaaggggt agaaatcctc ttgccgcctg
     8521 aggatagcgt gcagttcgcc caggtgaatc tggtggggga tgcttccatt gtcgaaagtg
     8581 cgctgtttgc gcaacagatc ttctctgtta agctttacca gcagctcctc ggtgccgtcc
     8641 attttttcca agatgggctt aataaatttg taaaattcct cctggcttgc tccgccgtca
     8701 atgtatccgg cgtagccatt tttagactga tcgaagaaaa tttccttgta cttctcaggc
     8761 agttgctgtc tgacaagggc cttcagcaaa gtcaagtctt ggtggtgctc atcatagcgc
     8821 ttgatcatac tagcgctcag cggagctttg gtgatctccg tgttcactcg cagaatatca
     8881 ctcagcagaa tggcgtctga caggttcttt gccgccaaaa aaaggtctgc gtactggtcg
     8941 ccgatctggg ccagcagatt gtcgagatca tcatcgtagg tgtctttgct cagttgaagc
     9001 ttggcatctt cggccaggtc gaagttagat ttaaagttgg gggtcagccc gagtgacagg
     9061 gcgataagat taccaaacag gccgttcttc ttctccccag ggagctgtgc gatgaggttt
     9121 tcgagccgcc gggatttgga cagcctagcg ctcaggattg ctttggcgtc aactccggat
     9181 gcgttgatcg ggttctcttc gaaaagctga ttgtaagtct gaaccagttg gataaagagt
     9241 ttgtcgacat cgctgttgtc tgggttcagg tccccctcga tgaggaagtg tccccgaaat
     9301 ttgatcatat gcgccagcgc gagatagatc aaccgcaagt cagccttatc agtactgtct
     9361 acaagcttct tcctcagatg atatatggtt gggtactttt catggtacgc cacctcgtcc
     9421 acgatattgc caaagattgg gtggcgctcg ttctttttat cctcctccac caaaaaggac
     9481 tcctccagcc tatggaagaa agagtcatcc accttagcca tctcattact aaagatctcc
     9541 tgcaggtagc agatccgatt ctttctgcgg gtatatctgc gccgtgctgt tcttttgagc
     9601 cgcgtggctt cggccgtctc cccggagtcg aacaggaggg cgccaatgag gttcttcttt
     9661 atgctgtggc gatcggtatt gcccagaact ttgaattttt tgctcggcac cttgtactcg
     9721 tccgtaatga cggcccaacc gacgctgttt gtgccgatat cgagcccaat ggagtacttc
     9781 ttgtccattt ttttccaagt tgattgtatg cttggtatag cttgaaatat tgtgcagaaa
     9841 aagaaacaag gaagaaaggg aacgagaaca atgacgagga aacaaaagat taataattgc
     9901 aggtctattt atacttgata gcaagacagc aaactttttt ttatttcaaa ttcaagtaac
     9961 tggaaggaag gccgtatacc gttgctcatt agagagtagt gtgcgtgaat gaaggaagga
    10021 aaaagtttcg tgtgcttcga gatacccctc atcagctctg gaacaacgac atctgttggt
    10081 gctgtctttg tcgttaattt tttcctttag tgtcttccat catttttttg tcattgcgga
    10141 tatggtgaga caacaacggg ggagagagaa aagaaaaaaa aagaaaagaa gttgcatgcg
    10201 cctattatta cttcaataga tggcaaatgg aaaaagggta gtgaaacttc gatatgatga
    10261 tggctatcaa gtctagggct acagtattag ttcgttatgt accaccatca atgaggcagt
    10321 gtaattggtg tagtcttgtt tagcccatta tgtcttgtct ggtatctgtt ctattgtata
    10381 tctcccctcc gccacctaca tgttagggag accaacgaag gtattatagg aatcccgatg
    10441 tatgggtttg gttgccagaa aagaggaagt ccatattgta cacccggaaa caacaaaagg
    10501 atgtcgacta tccactagac agaagtttgc gttccatact gtcagaggtg gcattttaca
    10561 taacaatagt gacaaaaaat aaaaaaagca ccgactcggt gccacttttt caagttgata
    10621 acggactagc cttattttaa cttgctattt ctagctctaa aacacatgct cgcttaacta
    10681 gtcaaagtcc cattcgccac ccgaaggtgt tgcccagccg gcgccagcga ggaggctggg
    10741 accatgccgg ccatctctcc cgggggcgag tcgaacgccc gatctcaaga tttcgtagtg
    10801 ataaattaca gtcttgcgcc ttaaaccaac ttggctaccg agagtattta attgttgaag
    10861 aaagagtata ctagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
    10921 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    10981 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    11041 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    11101 taatga
//
