LOCUS       sequence-231494-       10757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes yeast-optimized dCas9_VP64 synthetic transcription factor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10757)
  AUTHORS   Farzadfard F, Perli SD, Lu TK
  TITLE     Tunable and Multifunctional Eukaryotic Transcription Factors Based 
            on CRISPR/Cas.
  JOURNAL   ACS Synth Biol. 2013 Sep 11.
  PUBMED    23977949
REFERENCE   2  (bases 1 to 10757)
  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"; journalName: "ACS Synth 
            Biol. 2013 Sep 11."
COMMENT     Sequence Label: pTPGI_dCas9_VP64
FEATURES             Location/Qualifiers
     source          1..10757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        235..676
                     /label=TPGI promoter
                     /note="S. cerevisiae GAL1 promoter modified to contain two 
                     copies of the tet operator (Ellis et al., 2009)"
     protein_bind    235..352
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     protein_bind    560..578
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    581..599
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     647..670
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             716..736
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(746..769,770..772,773..3259,3260..3262,3263..4846)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4883..5032
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(5064..5082)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      5094..5283
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5355..5372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5526..6114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5606..5625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6285..7076,7077..7145))
                     /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     6908..6927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7146..7250)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7318..7336
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7374..7396)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7496..7515
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7536..7817
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7818..8492
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(8595..9050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8732..8753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8944..8963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(9128..9293)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     9214..9238
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(9437..10060)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     9968..9987
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(10078..10484)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacgttgcga
       61 agagcgacaa agattttgtt atcagcttta ttgctcaaag agacatgggt ggaagagatg
      121 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
      181 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggaattcga agtacggatt
      241 agaagccgcc gagcgggtga cagccctccg aaggaagact ctcctccgtg cgtcctcgtc
      301 ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa
      361 agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc
      421 cacaaacctt caaatgaacg aatcaaatta acaaccatag gatgataatg cgattacttt
      481 tttagcctta tttctggggt actgcagcag cgaagcgatg atttttgatc tattaacaga
      541 tatataaatg caaaaactgt ccctatcagt gatagagatc tccctatcag tgatagagat
      601 attacttctt attcaaatgt aataaaagta tcaacaaaaa attgttaata tacctctata
      661 ctttaacgtc aaggagaaaa accccggatc cacgattaaa agaatgtcta gagccccaaa
      721 gaagaagaga aaagttagac ccggggataa gaaatactct attggtttgg ctatcggtac
      781 aaactctgtt ggttgggctg ttattactga tgaatacaag gttccatcca agaagttcaa
      841 ggttttgggt aacactgata gacactccat caaaaagaac ttgattggtg ccttgttgtt
      901 cgattctggt gaaactgctg aagctactag attgaaaaga accgctagaa gaagatacac
      961 cagaagaaag aacagaatct gctacttgca agaaatcttc tccaacgaaa tggccaaggt
     1021 tgatgattca ttcttccaca gattggaaga atccttcttg gtcgaagaag ataagaagca
     1081 cgaaagacat ccaatcttcg gtaacatcgt tgatgaagtt gcttaccacg aaaagtaccc
     1141 aactatctac catttgagaa agaagttggt tgactctacc gataaggctg atttgagatt
     1201 gatctatttg gctttggccc acatgattaa gttcagaggt catttcttga tcgaaggtga
     1261 tttgaaccca gataactccg atgttgataa gttgttcatc caattagtcc aaacctacaa
     1321 tcaattattc gaagaaaacc caatcaacgc ctctggtgtt gatgctaaag ctattttgtc
     1381 tgccagattg tccaagtcca gaagattaga aaatttgatc gcccaattac caggtgaaaa
     1441 gaagaatggt ttgttcggta atttgattgc cttgtctttg ggtttgactc caaacttcaa
     1501 gtccaatttc gatttggctg aagatgccaa gttgcaatta tctaaggata cctacgatga
     1561 cgatttggat aacttgttgg ctcaaatcgg tgatcaatac gctgatttgt ttttggctgc
     1621 taagaacttg tccgatgcca ttttgttgtc cgatattttg agagtcaaca ccgaaattac
     1681 taaggctcca ttgtctgcct ctatgatcaa aagatacgat gaacaccacc aagacttgac
     1741 tttgttgaag gctttggtca gacaacaatt acctgaaaag tacaaagaaa ttttcttcga
     1801 tcaatccaag aacggttacg ccggttatat tgatggtggt gcttctcaag aagaatttta
     1861 caagttcatc aagccaatct tggaaaagat ggacggtact gaagaattat tggtcaagtt
     1921 gaacagagaa gatttgttga gaaagcaaag aaccttcgac aacggttcta ttccacatca
     1981 aattcacttg ggtgaattgc acgcaatttt gagaagacaa gaagattttt atccattctt
     2041 gaaggacaac agagaaaaga tcgaaaagat tctgaccttc agaatccctt actacgttgg
     2101 tccattggct agaggtaatt caagatttgc ctggatgact agaaagtccg aagaaactat
     2161 tactccttgg aacttcgaag aagttgtaga taagggtgct tctgcccaat cctttattga
     2221 aagaatgacc aacttcgaca agaacttgcc aaacgaaaag gttttgccaa agcactcttt
     2281 gttgtacgaa tacttcaccg tctacaacga attgactaag gttaagtacg tcaccgaagg
     2341 tatgagaaaa ccagcttttt tatccggtga acaaaagaag gctatcgtcg atttgttgtt
     2401 caagaccaac agaaaggtta ctgtcaagca attaaaagaa gattacttca agaaaatcga
     2461 atgcttcgac tccgttgaaa tttctggtgt cgaagataga ttcaatgcct ctttaggtac
     2521 ttaccatgac ttgttgaaaa tcatcaagga caaggatttc ttggacaacg aagaaaacga
     2581 agatattttg gaagatattg tcttgacatt gaccttgttt gaagatagag aaatgattga
     2641 agaaagattg aaaacctacg cccacttgtt cgatgataag gttatgaagc aattaaagag
     2701 aagaagatac actggttggg gtagattgtc cagaaaattg attaacggta tcagagacaa
     2761 gcaatccggt aagaccattt tggacttttt gaagtctgat ggtttcgcta acagaaactt
     2821 catgcaatta atccacgacg attccttgac tttcaaagaa gatatacaaa aggcccaagt
     2881 ctctggtcaa ggtgattctt tacatgaaca tatcgctaac ttggctggtt ctccagctat
     2941 taagaagggt attttacaaa ccgttaaggt cgttgacgaa ttggtcaaag ttatgggtag
     3001 acataagcca gaaaacatcg ttatcgaaat ggctagagaa aatcaaacca cccaaaaggg
     3061 tcaaaagaac tccagagaaa gaatgaagag aatcgaagaa ggtatcaaag aattgggttc
     3121 ccaaattttg aaagaacacc cagttgaaaa cacccaatta caaaacgaaa agttgtactt
     3181 gtactacttg caaaacggta gagatatgta cgttgaccaa gaattggaca tcaacagatt
     3241 gtctgattac gatgttgacg ctatcgttcc acaatctttt ttgaaggatg actccattga
     3301 caacaaggtc ttgactagat ccgataagaa tagaggtaag tccgataacg ttccatctga
     3361 agaagtcgtt aagaaaatga agaactattg gagacaatta ttgaacgcca agttgatcac
     3421 ccaaagaaag tttgacaatt tgaccaaggc tgaaagaggt ggtttgtctg aattggataa
     3481 ggcaggtttc atcaaaagac aattagtaga aaccagacaa atcaccaagc acgttgctca
     3541 aattttggat agtagaatga acactaagta cgacgaaaac gacaaattga tcagagaagt
     3601 taaggtcatt accttgaagt ccaagttggt ttccgatttc agaaaggact tccaattcta
     3661 caaggtcaga gaaatcaaca actaccatca tgcacatgat gcttacttga atgctgttgt
     3721 tggtactgcc ttgattaaga agtatccaaa gttggaatcc gaatttgtct acggtgatta
     3781 caaggtttac gacgttagaa agatgatcgc caagtccgaa caagaaattg gtaaagctac
     3841 tgccaaatac ttcttctact ccaatattat gaatttcttt aagaccgaaa tcactttggc
     3901 caacggtgaa attagaaaaa gaccattgat tgaaactaat ggtgaaacag gtgaaatcgt
     3961 ttgggataag ggtagagatt ttgccactgt tagaaaggta ttgtccatgc cacaagtaaa
     4021 catcgtcaaa aagaccgaag ttcaaactgg tggtttctcc aaagaatcca ttttgcctaa
     4081 gagaaactcc gataagttga tcgctagaaa aaaagactgg gacccaaaaa agtacggtgg
     4141 ttttgattct ccaactgttg cttactctgt tttggttgtt gctaaggtcg aaaagggtaa
     4201 gagtaagaag ttgaagtccg tcaaagaatt attaggtatc actatcatgg aaagatcctc
     4261 attcgaaaag aatcctatcg actttttgga agccaagggt tacaaagaag tcaagaagga
     4321 cttgatcatt aagttgccaa agtacagttt gttcgaattg gaaaatggta gaaagagaat
     4381 gttggcttct gccggtgaat tacaaaaggg taatgaattg gctttgccat ccaagtacgt
     4441 taatttctta tacttggcct cccactacga aaaattgaaa ggttctcctg aagataacga
     4501 acaaaagcaa ttatttgtcg aacaacacaa gcactacttg gacgaaatca ttgaacaaat
     4561 ttccgaattt tccaaaagag tcattttggc tgacgccaat ttggacaaag ttttgtcagc
     4621 ttacaacaag cacagagata agccaattag agaacaagct gaaaacatca ttcacttgtt
     4681 cactttgact aacttgggtg ctccagctgc ttttaagtat ttcgatacca ctatcgacag
     4741 aaagagatac acctctacca aagaagtttt ggacgctact ttgatccacc aatctattac
     4801 tggtttgtac gaaactagaa tcgacttgtc tcaattaggt ggtgatggtt ctggtagatc
     4861 tggagtcgac ggtggaggtt ctgacgcttt ggacgacttc gacttggata tgctgggttc
     4921 tgatgcgcta gatgactttg acctcgacat gcttggaagt gacgccttag atgattttga
     4981 cctggatatg cttggatcag acgctctgga cgatttcgac ttagacatgc tttcctagga
     5041 actcgagtca tgtaattagt tatgtcacgc ttacattcac gcccttcccc cacatccgct
     5101 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
     5161 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     5221 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     5281 aagatccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5341 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5401 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5461 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5521 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5581 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5641 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5701 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5761 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5821 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5881 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5941 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6001 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6061 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6121 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6181 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6241 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6301 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6361 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6421 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6481 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6541 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6601 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6661 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6721 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6781 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6841 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6901 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6961 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7021 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7081 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7141 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7201 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7261 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7321 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     7381 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     7441 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     7501 tcagagcaga ttgtactgag agtgcaccat agatcaacga cattactata tatataatat
     7561 aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta tgacgccaga
     7621 tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg tacaatcttg
     7681 atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa agaaaaggag
     7741 agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg attaagcaca
     7801 caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt ccattggtga
     7861 aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc gatgctgact
     7921 tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt attgcaagga
     7981 aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa tacttggttg
     8041 gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat tacggcattg
     8101 atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc
     8161 cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac
     8221 agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt
     8281 tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa agcttacatt
     8341 ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg
     8401 ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa
     8461 tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt
     8521 attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     8581 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     8641 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     8701 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     8761 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     8821 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     8881 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     8941 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     9001 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     9061 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccac
     9121 tgaattggag cgacctcatg ctatacctga gaaagcaacc tgacctacag gaaagagtta
     9181 ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt gtatacactt
     9241 atttttttta taacttattt aataataaaa atcataaatc ataagaaatt cgcttattta
     9301 gaagtgtcaa caacgtatct accaacgatt tgaccctttt ccatcttttc gtaaatttct
     9361 ggcaaggtag acaagccgac aaccttgatt ggagacttga ccaaacctct ggcgaagaat
     9421 tgttaattaa gagctcttaa gacccacttt cacatttaag ttgtttttct aatccgcata
     9481 tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga
     9541 tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag
     9601 cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga
     9661 gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt
     9721 cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc
     9781 gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc
     9841 tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg
     9901 cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta
     9961 gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta
    10021 atgcgctgtt aatcacttta cttttatcta atctagacat tttaataact agtaaaactt
    10081 agattagatt gctatgcttt ctttctaatg agcaagaagt aaaaaaagtt gtaatagaac
    10141 aagaaaaatg aaactgaaac ttgagaaatt gaagaccgtt tattaactta aatatcaatg
    10201 ggaggtcatc gaaagagaaa aaaatcaaaa aaaaaatttt caagaaaaag aaacgtgata
    10261 aaaattttta ttgccttttt cgacgaagaa aaagaaacga ggcggtctct tttttctttt
    10321 ccaaaccttt agtacgggta attaacgaca ccctagagga agaaagaggg gaaatttagt
    10381 atgctgtgct tgggtgtttt gaagtggtac ggcgatgcgc ggagtccgag aaaatctgga
    10441 agagtaaaaa aggagtagaa acattttgaa gctatggtgt ggcggccgca tcttgactga
    10501 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
    10561 actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
    10621 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
    10681 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
    10741 tttgatgtta gcagaat
//
