LOCUS       sequence-352558-       10933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10933)
  AUTHORS   Kwak S, Mahmud B, Dantas G
  TITLE     A Tunable and Expandable Transactivation System in Probiotic Yeast 
            Saccharomyces boulardii.
  JOURNAL   ACS Synth Biol. 2022 Jan 21;11(1):508-514. doi: 
            10.1021/acssynbio.1c00384. Epub 2021 Dec 23.
  PUBMED    34939781
REFERENCE   2  (bases 1 to 10933)
  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: 
            "10.1021/acssynbio.1c00384"; journalName: "ACS Synth Biol"; date: 
            "2022-01-21- 21"; volume: "11"; issue: "1"; pages: "508-514"
COMMENT     Sequence Label: pRS42H_PTEF1_dCas9_TCYC1
FEATURES             Location/Qualifiers
     source          1..10933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     123..142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     376..393
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    510..531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(546..551,552..569,570..576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    584..600
                     /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     589..611
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     608..624
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     608..624
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     643..663
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        645..663
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        681..1079
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             join(1095..1121,1122..1124,1125..3608,3609..3614,3615..3680,
                     3681..3683,3684..5198)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5211..5231
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5247..5494
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5512..5531)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5513..5531)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5541..5558)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5541..5557)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5550..5572)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5698..6153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5785..5804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5995..6016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6438..7466)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVPADSGNRRPSTRPRA
                     KE"
     promoter        complement(7467..7810)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(7824..7843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7943..7965
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8003..8021)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      8062..9404
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    8394..8441
                     /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        9431..9535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9536..9604,9605..10396)
                     /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(9754..9773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10567..10933,1..222)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
       61 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      301 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      361 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      421 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      481 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      541 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      601 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      661 agggaacaaa agctggagct catagcttca aaatgtttct actccttttt tactcttcca
      721 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
      781 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
      841 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
      901 acgtttcttt ttcttgaaaa tttttttttt gatttttttc tctttcgatg acctcccatt
      961 gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc
     1021 tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagt
     1081 tttctagaac tagtatggac aagaagtact ccattgggct cgctatcggc acaaacagcg
     1141 tcggctgggc cgtcattacg gacgagtaca aggtgccgag caaaaaattc aaagttctgg
     1201 gcaataccga tcgccacagc ataaagaaga acctcattgg cgccctcctg ttcgactccg
     1261 gggagacggc cgaagccacg cggctcaaaa gaacagcacg gcgcagatat acccgcagaa
     1321 agaatcggat ctgctacctg caggagatct ttagtaatga gatggctaag gtggatgact
     1381 ctttcttcca taggctggag gagtcctttt tggtggagga ggataaaaag cacgagcgcc
     1441 acccaatctt tggcaatatc gtggacgagg tggcgtacca tgaaaagtac ccaaccatat
     1501 atcatctgag gaagaagctt gtagacagta ctgataaggc tgacttgcgg ttgatctatc
     1561 tcgcgctggc gcatatgatc aaatttcggg gacacttcct catcgagggg gacctgaacc
     1621 cagacaacag cgatgtcgat aaactcttta tccaactggt tcagacttac aatcagcttt
     1681 tcgaagagaa cccgatcaac gcatccggag ttgacgccaa agcaatcctg agcgctaggc
     1741 tgtccaaatc ccggcggctc gaaaacctca tcgcacagct ccctggggag aagaagaacg
     1801 gcctgtttgg taatcttatc gccctgtcac tcgggctgac ccccaacttt aaatctaact
     1861 tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga cacctacgat gatgatctcg
     1921 acaatctgct ggcccagatc ggcgaccagt acgcagacct ttttttggcg gcaaagaacc
     1981 tgtcagacgc cattctgctg agtgatattc tgcgagtgaa cacggagatc accaaagctc
     2041 cgctgagcgc tagtatgatc aagcgctatg atgagcacca ccaagacttg actttgctga
     2101 aggcccttgt cagacagcaa ctgcctgaga agtacaagga aattttcttc gatcagtcta
     2161 aaaatggcta cgccggatac attgacggcg gagcaagcca ggaggaattt tacaaattta
     2221 ttaagcccat cttggaaaaa atggacggca ccgaggagct gctggtaaag cttaacagag
     2281 aagatctgtt gcgcaaacag cgcactttcg acaatggaag catcccccac cagattcacc
     2341 tgggcgaact gcacgctatc ctcaggcggc aagaggattt ctaccccttt ttgaaagata
     2401 acagggaaaa gattgagaaa atcctcacat ttcggatacc ctactatgta ggccccctcg
     2461 cccggggaaa ttccagattc gcgtggatga ctcgcaaatc agaagagacc atcactccct
     2521 ggaacttcga ggaagtcgtg gataaggggg cctctgccca gtccttcatc gaaaggatga
     2581 ctaactttga taaaaatctg cctaacgaaa aggtgcttcc taaacactct ctgctgtacg
     2641 agtacttcac agtttataac gagctcacca aggtcaaata cgtcacagaa gggatgagaa
     2701 agccagcatt cctgtctgga gagcagaaga aagctatcgt ggacctcctc ttcaagacga
     2761 accggaaagt taccgtgaaa cagctcaaag aagactattt caaaaagatt gaatgtttcg
     2821 actctgttga aatcagcgga gtggaggatc gcttcaacgc atccctggga acgtatcacg
     2881 atctcctgaa aatcattaaa gacaaggact tcctggacaa tgaggagaac gaggacattc
     2941 ttgaggacat tgtcctcacc cttacgttgt ttgaagatag ggagatgatt gaagaacgct
     3001 tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa acagctcaag aggcgccgat
     3061 atacaggatg ggggcggctg tcaagaaaac tgatcaatgg gatccgagac aagcagagtg
     3121 gaaagacaat cctggatttt cttaagtccg atggatttgc caaccggaac ttcatgcagt
     3181 tgatccatga tgactctctc acctttaagg aggacatcca gaaagcacaa gtttctggcc
     3241 agggggacag tcttcacgag cacatcgcta atcttgcagg tagcccagct atcaaaaagg
     3301 gaatactgca gaccgttaag gtcgtggatg aactcgtcaa agtaatggga aggcataagc
     3361 ccgagaatat cgttatcgag atggcccgag agaaccaaac tacccagaag ggacagaaga
     3421 acagtaggga aaggatgaag aggattgaag agggtataaa agaactgggg tcccaaatcc
     3481 ttaaggaaca cccagttgaa aacacccagc ttcagaatga gaagctctac ctgtactacc
     3541 tgcagaacgg cagggacatg tacgtggatc aggaactgga catcaatcgg ctctccgact
     3601 acgacgtggc tgctatcgtg ccccagtctt ttctcaaaga tgattctatt gataataaag
     3661 tgttgacaag atccgataaa gctagaggga agagtgataa cgtcccctca gaagaagttg
     3721 tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc caaactgatc acacaacgga
     3781 agttcgataa tctgactaag gctgaacgag gtggcctgtc tgagttggat aaagccggct
     3841 tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa gcacgtggcc caaattctcg
     3901 attcacgcat gaacaccaag tacgatgaaa atgacaaact gattcgagag gtgaaagtta
     3961 ttactctgaa gtctaagctg gtctcagatt tcagaaagga ctttcagttt tataaggtga
     4021 gagagatcaa caattaccac catgcgcatg atgcctacct gaatgcagtg gtaggcactg
     4081 cacttatcaa aaaatatccc aagcttgaat ctgaatttgt ttacggagac tataaagtgt
     4141 acgatgttag gaaaatgatc gcaaagtctg agcaggaaat aggcaaggcc accgctaagt
     4201 acttctttta cagcaatatt atgaattttt tcaagaccga gattacactg gccaatggag
     4261 agattcggaa gcgaccactt atcgaaacaa acggagaaac aggagaaatc gtgtgggaca
     4321 agggtaggga tttcgcgaca gtccggaagg tcctgtccat gccgcaggtg aacatcgtta
     4381 aaaagaccga agtacagacc ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca
     4441 gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa gaaatacggc ggattcgatt
     4501 ctcctacagt cgcttacagt gtactggttg tggccaaagt ggagaaaggg aagtctaaaa
     4561 aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat ggagcgatca agcttcgaaa
     4621 aaaaccccat cgactttctc gaggcgaaag gatataaaga ggtcaaaaaa gacctcatca
     4681 ttaagcttcc caagtactct ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta
     4741 gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc ctctaaatac gttaatttct
     4801 tgtatctggc cagccactat gaaaagctca aagggtctcc cgaagataat gagcagaagc
     4861 agctgttcgt ggaacaacac aaacactacc ttgatgagat catcgagcaa ataagcgaat
     4921 tctccaaaag agtgatcctc gccgacgcta acctcgataa ggtgctttct gcttacaata
     4981 agcacaggga taagcccatc agggagcagg cagaaaacat tatccacttg tttactctga
     5041 ccaacttggg cgcgcctgca gccttcaagt acttcgacac caccatagac agaaagcggt
     5101 acacctctac aaaggaggtc ctggacgcca cactgattca tcagtcaatt acggggctct
     5161 atgaaacaag aatcgacctc tctcagctcg gtggagacag cagggctgac cccaagaaga
     5221 agaggaaggt gtaagtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg
     5281 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     5341 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     5401 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     5461 gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt
     5521 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5581 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5641 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     5701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6121 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     6181 tctgtgcggt atttcacacc gccgtcccaa accttctcaa gcaaggtttt cagtataatg
     6241 ttacatgcgt acacgcgtct gtacagaaaa aaaagaaaaa tttgaaatat aaataacgtt
     6301 cttaatacta acataactat aaaaaaataa atagggacct agacttcagg ttgtctaact
     6361 ccttcctttt cggttagagc ggatgtgggg ggagggcgtg aatgtaagcg tgacataact
     6421 aattacatga ctcgagatta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca
     6481 ctatcggcgg gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt
     6541 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc
     6601 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg
     6661 cggagcatat acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag
     6721 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc
     6781 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca
     6841 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg
     6901 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg
     6961 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca
     7021 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg
     7081 ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc
     7141 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag
     7201 gtcaggctct cgctgaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat
     7261 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
     7321 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
     7381 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
     7441 gcggtgagtt caggcttttt acccatggtt gtttatgttc ggatgtgatg tgagaactgt
     7501 atcctagcaa gattttaaaa ggaagtatat gaaagaagaa cctcagtggc aaatcctaac
     7561 cttttatatt tctctacagg ggcgcggcgt ggggacaatt caacgcgtct gtgaggggag
     7621 cgtttccctg ctcgcaggtc cgcagcgagg agccgtaatt tttgcttcgc gccgtgcggc
     7681 catcaaaatg tatggatgca aatgattata catggggatg tatgggctaa atgtacgggc
     7741 gacagtcaca tcatgcccct gagctgcgca cgtcaagact gtcaaggagg gtattctggg
     7801 cctccatgtc gctgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     7861 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     7921 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     7981 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     8041 ataataatgg tttcttagta tgatccaata tcaaaggaaa tgatagcatt gaaggatgag
     8101 actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc tgaaggaagc
     8161 atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta cctttcatcc
     8221 tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat gccgttcttc
     8281 tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt
     8341 atgtgcgcag ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc
     8401 ctattccgaa gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaaaacca
     8461 aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac
     8521 taaaatattg cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat
     8581 ctctgtgcta tatccctata taacctaccc atccaccttt cgctccttga acttgcatct
     8641 aaactcgacc tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt
     8701 gtagtaagaa ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat
     8761 acgtagtata tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat
     8821 catcaacgct atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata
     8881 atcggggatg cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg
     8941 cgggaagtgg agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt
     9001 ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca
     9061 aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt
     9121 tttgtagaac aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca
     9181 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
     9241 gcatttttgt tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg
     9301 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     9361 gcgttgcatt tttgttctac aaaatgaagc acagatgctt cgttcaggtg gcacttttcg
     9421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     9481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     9541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     9601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     9661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     9721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     9781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     9841 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     9901 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     9961 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    10021 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    10081 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    10141 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    10201 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    10261 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    10321 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    10381 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    10441 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    10501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    10561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    10621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    10681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    10741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    10801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    10861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    10921 aacgacctac acc
//
