LOCUS       sequence-193432-       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 SAP155c sequence 
            ATGAAAGACAACTATAGGGC in yeast chromosome 6.
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-SAP155c
FEATURES             Location/Qualifiers
     source          1..11106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(73..288)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(312..331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(491..509)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      550..1892
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    882..929
                     /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        1919..2023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2024..2092,2093..2884)
                     /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(2242..2261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3055..3643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3544..3563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3797..3814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3931..3952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3967..3972,3973..3990,3991..3997)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4005..4021
                     /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     4010..4032
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4029..4045
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4029..4045
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4064..4084
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4066..4084
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        4311..4386
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4472..5176
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5186..9289
                     /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             9302..9322
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9338..9585
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9603..9622)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9604..9622)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9632..9649)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9632..9648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9641..9663)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9789..10244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9876..9895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10086..10107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(10375..11106,1..72))
                     /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 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
       61 agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa
      121 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
      181 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
      241 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg
      301 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      361 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      421 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
      481 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
      541 tcttagtatg atccaatatc aaaggaaatg atagcattga aggatgagac taatccaatt
      601 gaggagtggc agcatataga acagctaaag ggtagtgctg aaggaagcat acgatacccc
      661 gcatggaatg ggataatatc acaggaggta ctagactacc tttcatccta cataaataga
      721 cgcatataag tacgcattta agcataaaca cgcactatgc cgttcttctc atgtatatat
      781 atatacaggc aacacgcaga tataggtgcg acgtgaacag tgagctgtat gtgcgcagct
      841 cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt tgaagttcct attccgaagt
      901 tcctattctc tagaaagtat aggaacttca gagcgctttt gaaaaccaaa agcgctctga
      961 agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta aaatattgcg
     1021 aataccgctt ccacaaacat tgctcaaaag tatctctttg ctatatatct ctgtgctata
     1081 tccctatata acctacccat ccacctttcg ctccttgaac ttgcatctaa actcgacctc
     1141 tacatttttt atgtttatct ctagtattac tctttagaca aaaaaattgt agtaagaact
     1201 attcatagag tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac gtagtatata
     1261 gagacaaaat agaagaaacc gttcataatt ttctgaccaa tgaagaatca tcaacgctat
     1321 cactttctgt tcacaaagta tgcgcaatcc acatcggtat agaatataat cggggatgcc
     1381 tttatcttga aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag
     1441 tcaggctttt tttatggaag agaaaataga caccaaagta gccttcttct aaccttaacg
     1501 gacctacagt gcaaaaagtt atcaagagac tgcattatag agcgcacaaa ggagaaaaaa
     1561 agtaatctaa gatgctttgt tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa
     1621 aaaagaagta tagattcttt gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt
     1681 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt
     1741 tacaaaaatg aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt
     1801 tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt
     1861 tgttctacaa aatgaagcac agatgcttcg ttcaggtggc acttttcggg gaaatgtgcg
     1921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2041 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2161 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2221 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2401 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2581 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2641 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2881 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2941 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3001 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3061 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3181 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3721 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3781 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3841 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     3901 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     3961 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     4021 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     4081 aagggaacaa aagctggagc tagtatactc tttcttcaac aattaaatac tctcggtagc
     4141 caagttggtt taaggcgcaa gactgtaatt tatcactacg aaatcttgag atcgggcgtt
     4201 cgactcgccc ccgggagaga tggccggcat ggtcccagcc tcctcgctgg cgccggctgg
     4261 gcaacacctt cgggtggcga atgggacttt atgaaagaca actatagggc gttttagagc
     4321 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     4381 cggtgctttt tttatttttt gtcactattg ttatgtaaaa tgccacctct gacagtatgg
     4441 aacgcaaact tctgtctagt ggatagtcga catccttttg ttgtttccgg gtgtacaata
     4501 tggacttcct cttttctggc aaccaaaccc atacatcggg attcctataa taccttcgtt
     4561 ggtctcccta acatgtaggt ggcggagggg agatatacaa tagaacagat accagacaag
     4621 acataatggg ctaaacaaga ctacaccaat tacactgcct cattgatggt ggtacataac
     4681 gaactaatac tgtagcccta gacttgatag ccatcatcat atcgaagttt cactaccctt
     4741 tttccatttg ccatctattg aagtaataat aggcgcatgc aacttctttt cttttttttt
     4801 cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa atgatggaag
     4861 acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg ttccagagct
     4921 gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg cacactactc
     4981 tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa taaaaaaaag
     5041 tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt gtttcctcgt
     5101 cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc aagctatacc
     5161 aagcatacaa tcaacttgga aaaaaatgga caagaagtac tccattgggc tcgatatcgg
     5221 cacaaacagc gtcggttggg ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt
     5281 caaagttctg ggcaataccg atcgccacag cataaagaag aacctcattg gcgccctcct
     5341 gttcgactcc ggggagacgg ccgaagccac gcggctcaaa agaacagcac ggcgcagata
     5401 tacccgcaga aagaatcgga tctgctacct gcaggagatc tttagtaatg agatggctaa
     5461 ggtggatgac tctttcttcc ataggctgga ggagtccttt ttggtggagg aggataaaaa
     5521 gaacgagcgc cacccaatct ttggcaatat cgtggacgag gtggcgtacc atgaaaagta
     5581 cccaaccata tatcatctga ggaagaagct tgtagacagt actgataagg ctgacttgcg
     5641 gttgatctat ctcgcgctgg cgcatatgat caaatttcgg ggacacttcc tcatcgaggg
     5701 ggacctgaac ccagacaaca gcgatgtcga caaactcttt atccaactgg ttcagactta
     5761 caatcagctt ttcgaagaga acccgatcaa cgcatccgga gttgacgcca aagcaatcct
     5821 gagcgctagg ctgtccaaat cccggcggct cgaaaacctc atcgcacagc tccctgggga
     5881 gaagaagaac ggcctgtttg gtaatcttat cgccctgtca ctcgggctga cccccaactt
     5941 taaatctaac ttcgacctgg ccgaagatgc caagcttcaa ctgagcaaag acacctacga
     6001 tgatgatctc gacaatctgc tggcccagat cggcgaccag tacgcagacc tttttttggc
     6061 ggcaaagaac ctgtcagacg ccattctgct gagtgatatt ctgcgagtga acacggagat
     6121 caccaaagct ccgctgagcg ctagtatgat caagcgctat gatgagcacc accaagactt
     6181 gactttgctg aaggcccttg tcagacagca actgcctgag aagtacaagg aaattttctt
     6241 cgatcagtct aaaaatggct acgccggata cattgacggc ggagcaagcc aggaggaatt
     6301 ttacaaattt attaagccca tcttggaaaa aatggacggc accgaggagc tgctggtaaa
     6361 gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc gacaatggaa gcatccccca
     6421 ccagattcac ctgggcgaac tgcacgctat cctcaggcgg caagaggatt tctacccctt
     6481 tttgaaagat aacagggaaa agattgagaa aatcctcaca tttcggatac cctactatgt
     6541 aggccccctc gcccggggaa attccagatt cgcgtggatg actcgcaaat cagaagagac
     6601 catcactccc tggaacttcg aggaagtcgt ggataagggg gcctctgccc agtccttcat
     6661 cgaaaggatg actaactttg ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc
     6721 tctgctgtac gagtacttca cagtttataa cgagctcacc aaggtcaaat acgtcacaga
     6781 agggatgaga aagccagcat tcctgtctgg agagcagaag aaagctatcg tggacctcct
     6841 cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa gaagactatt tcaaaaagat
     6901 tgaatgtttc gactctgttg aaatcagcgg agtggaggat cgcttcaacg catccctggg
     6961 aacgtatcac gatctcctga aaatcattaa agacaaggac ttcctggaca atgaggagaa
     7021 cgaggacatt cttgaggaca ttgtcctcac ccttacgttg tttgaagata gggagatgat
     7081 tgaagaacgc ttgaaaactt acgctcatct cttcgacgac aaagtcatga aacagctcaa
     7141 gaggcgccga tatacaggat gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga
     7201 caagcagagt ggaaagacaa tcctggattt tcttaagtcc gatggatttg ccaaccggaa
     7261 cttcatgcag ttgatccatg atgactctct cacctttaag gaggacatcc agaaagcaca
     7321 agtttctggc cagggggaca gtcttcacga gcacatcgct aatcttgcag gtagcccagc
     7381 tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat gaactcgtca aagtaatggg
     7441 aaggcataag cccgagaata tcgttatcga gatggcccga gagaaccaaa ctacccagaa
     7501 gggacagaag aacagtaggg aaaggatgaa gaggattgaa gagggtataa aagaactggg
     7561 gtcccaaatc cttaaggaac acccagttga aaacacccag cttcagaatg agaagctcta
     7621 cctgtactac ctgcagaacg gcagggacat gtacgtggat caggaactgg acatcaatcg
     7681 gctctccgac tacgacgtgg atcatatcgt gccccagtct tttctcaaag atgattctat
     7741 tgataataaa gtgttgacaa gatccgataa aaatagaggg aagagtgata acgtcccctc
     7801 agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat
     7861 cacacaacgg aagttcgata atctgactaa ggctgaacga ggtggcctgt ctgagttgga
     7921 taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc cagatcacca agcacgtggc
     7981 ccaaattctc gattcacgca tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga
     8041 ggtgaaagtt attactctga agtctaagct ggtctcagat ttcagaaagg actttcagtt
     8101 ttataaggtg agagagatca acaattacca ccatgcgcat gatgcctacc tgaatgcagt
     8161 ggtaggcact gcacttatca aaaaatatcc caagcttgaa tctgaatttg tttacggaga
     8221 ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc
     8281 caccgctaag tacttctttt acagcaatat tatgaatttt ttcaagaccg agattacact
     8341 ggccaatgga gagattcgga agcgaccact tatcgaaaca aacggagaaa caggagaaat
     8401 cgtgtgggac aagggtaggg atttcgcgac agtccggaag gtcctgtcca tgccgcaggt
     8461 gaacatcgtt aaaaagaccg aagtacagac cggaggcttc tccaaggaaa gtatcctccc
     8521 gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat tgggacccca agaaatacgg
     8581 cggattcgat tctcctacag tcgcttacag tgtactggtt gtggccaaag tggagaaagg
     8641 gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc atcacaatca tggagcgatc
     8701 aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa
     8761 agacctcatc attaagcttc ccaagtactc tctctttgag cttgaaaacg gccggaaacg
     8821 aatgctcgct agtgcgggcg agctgcagaa aggtaacgag ctggcactgc cctctaaata
     8881 cgttaatttc ttgtatctgg ccagccacta tgaaaagctc aaagggtctc ccgaagataa
     8941 tgagcagaag cagctgttcg tggaacaaca caaacactac cttgatgaga tcatcgagca
     9001 aataagcgaa ttctccaaaa gagtgatcct cgccgacgct aacctcgata aggtgctttc
     9061 tgcttacaat aagcacaggg ataagcccat cagggagcag gcagaaaaca ttatctactt
     9121 gtttactctg accaacttgg gcgcgcctgc agccttcaag tacttcgaca ccaccataga
     9181 cagaaagcgg tacacctcta caaaggaggt cctggacgcc acactgattc atcagtcaat
     9241 tacggggctc tatgaaacaa gaatcgacct ctctcagctc ggtggagaca gcagggctga
     9301 ccccaagaag aagaggaagg tgtgatctct tctcgagtca tgtaattagt tatgtcacgc
     9361 ttacgttcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     9421 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     9481 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     9541 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt
     9601 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     9661 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     9721 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     9781 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     9841 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     9901 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     9961 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
    10021 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
    10081 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
    10141 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
    10201 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
    10261 ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat
    10321 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
    10381 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
    10441 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
    10501 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
    10561 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
    10621 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
    10681 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
    10741 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
    10801 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
    10861 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
    10921 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
    10981 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
    11041 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
    11101 gtgcat
//
