LOCUS       sequence-109392-       11468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAG414 series plasmid with GPD promoter driving expression of 
            dCas9-VPR; cerevisiae vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11468)
  AUTHORS   Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin 
            S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan D, Braff JL, 
            Davidsohn N, Housden BE, Perrimon N, Weiss R, Aach J, Collins JJ, 
            Church GM
  TITLE     Highly efficient Cas9-mediated transcriptional programming.
  JOURNAL   Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.
  PUBMED    25730490
REFERENCE   2  (bases 1 to 11468)
  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: "Nat 
            Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312."
COMMENT     Sequence Label: pAG414GPD-dCas9-VPR
FEATURES             Location/Qualifiers
     source          1..11468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2804..3448
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3342..3365
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3454..3470
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3454..3470
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3478..3502
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3508..3534,3535..3537,3538..6021,6022..6027,6028..6093,
                     6094..6096,6097..7611)
                     /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             7624..7644
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7669..7818
                     /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"
     CDS             7843..7863
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8293..8649
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             8668..9237
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     protein_bind    complement(9242..9266)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9299..9315)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9300..9316)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      9318..9565
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9334..9352)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(9583..9602)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9584..9602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9612..9629)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9612..9628)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9621..9643)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9773..10228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9860..9879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10070..10091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(10327..11001)
                     /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"
     promoter        complement(11002..11282)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(11299..11318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11418..11440
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcttcgagt ttatcattat caatactcgc
     2821 catttcaaag aatacgtaaa taattaatag tagtgatttt cctaacttta tttagtcaaa
     2881 aaattagcct tttaattctg ctgtaacccg tacatgccca aaataggggg cgggttacac
     2941 agaatatata acaccgaagg tgcctgggtg aacaggttta ttcctggcat ccactaaata
     3001 taatggagcc cgctttttaa gctggcatcc agaaaaaaaa agaatcccag caccaaaata
     3061 ttgttttctt caccaaccat cagttcatag gtccattctc ttagcgcaac tacagagaac
     3121 aggggcacaa acaggcaaaa aacgggcaca acctcaatgg agtgatgcaa cctgcctgga
     3181 gtaaatgatg acacaaggca attgacccac gcatgtatct atctcatttt cttacacctt
     3241 ctattacctt ctgctctctc tgatttggaa aaagctgaaa aaaaaggttg aaaccagttc
     3301 cctgaaatta ttcccctact tgactaataa gtatataaag acggtaggta ttgattgtaa
     3361 ttctgtaaat ctatttctta aacttcttaa attctacttt tatagttagt ctttttttta
     3421 gttttaaaac accagaactt agtttcgacg gattctagaa ctagtggatc ccccatcaca
     3481 agtttgtaca aaaaagcagg ctacaaaatg gacaagaagt actccattgg gctcgctatc
     3541 ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa
     3601 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc
     3661 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga
     3721 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct
     3781 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa
     3841 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag
     3901 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg
     3961 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag
     4021 ggggacctga acccagacaa cagcgatgtc gataaactct ttatccaact ggttcagact
     4081 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc
     4141 ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg
     4201 gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac
     4261 tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac
     4321 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg
     4381 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag
     4441 atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac
     4501 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc
     4561 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa
     4621 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta
     4681 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc
     4741 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc
     4801 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat
     4861 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag
     4921 accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc
     4981 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac
     5041 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca
     5101 gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc
     5161 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag
     5221 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg
     5281 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag
     5341 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg
     5401 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc
     5461 aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga
     5521 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg
     5581 aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca
     5641 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca
     5701 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg
     5761 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag
     5821 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg
     5881 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc
     5941 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat
     6001 cggctctccg actacgacgt ggctgctatc gtgccccagt cttttctcaa agatgattct
     6061 attgataata aagtgttgac aagatccgat aaagctagag ggaagagtga taacgtcccc
     6121 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg
     6181 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg
     6241 gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg
     6301 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga
     6361 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag
     6421 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca
     6481 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga
     6541 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag
     6601 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca
     6661 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa
     6721 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag
     6781 gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc
     6841 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac
     6901 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa
     6961 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga
     7021 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa
     7081 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa
     7141 cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa
     7201 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat
     7261 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag
     7321 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt
     7381 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac
     7441 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata
     7501 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca
     7561 attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct
     7621 gaccccaaga agaagaggaa ggtggaggcc agcggttccg gacgggctga cgcattggac
     7681 gattttgatc tggatatgct gggaagtgac gccctcgatg attttgacct tgacatgctt
     7741 ggttcggatg cccttgatga ctttgacctc gacatgctcg gcagtgacgc ccttgatgat
     7801 ttcgacctgg acatgctgat taactctaga agttccggat ctccgaaaaa gaaacgcaaa
     7861 gttggtagcc agtacctgcc cgacaccgac gaccggcacc ggatcgagga aaagcggaag
     7921 cggacctacg agacattcaa gagcatcatg aagaagtccc ccttcagcgg ccccaccgac
     7981 cctagacctc cacctagaag aatcgccgtg cccagcagat ccagcgccag cgtgccaaaa
     8041 cctgcccccc agccttaccc cttcaccagc agcctgagca ccatcaacta cgacgagttc
     8101 cctaccatgg tgttccccag cggccagatc tctcaggcct ctgctctggc tccagcccct
     8161 cctcaggtgc tgcctcaggc tcctgctcct gcaccagctc cagccatggt gtctgcactg
     8221 gctcaggcac cagcacccgt gcctgtgctg gctcctggac ctccacaggc tgtggctcca
     8281 ccagccccta aacctacaca ggccggcgag ggcacactgt ctgaagctct gctgcagctg
     8341 cagttcgacg acgaggatct gggagccctg ctgggaaaca gcaccgatcc tgccgtgttc
     8401 accgacctgg ccagcgtgga caacagcgag ttccagcagc tgctgaacca gggcatccct
     8461 gtggcccctc acaccaccga gcccatgctg atggaatacc ccgaggccat cacccggctc
     8521 gtgacaggcg ctcagaggcc tcctgatcca gctcctgccc ctctgggagc accaggcctg
     8581 cctaatggac tgctgtctgg cgacgaggac ttcagctcta tcgccgacat ggacttctcc
     8641 gcactgctgg gtagcggatc gggatctcgg gattccaggg aagggatgtt tttgccgaag
     8701 cctgaggccg gctccgctat tagtgacgtg tttgagggcc gcgaggtgtg ccagccaaaa
     8761 cgaatccggc catttcatcc tccaggaagt ccatgggcca accgcccact ccccgccagc
     8821 ctcgcaccaa caccaaccgg tccagtacat gagccagtcg ggtcactgac cccggcacca
     8881 gtccctcagc cactggatcc agcgcccgca gtgactcccg aggccagtca cctgttggag
     8941 gatcccgatg aagagacgag ccaggctgtc aaagcccttc gggagatggc cgatactgtg
     9001 attccccaga aggaagaggc tgcaatctgt ggccaaatgg acctttccca tccgccccca
     9061 aggggccatc tggatgagct gacaaccaca cttgagtcca tgaccgagga tctgaacctg
     9121 gactcacccc tgaccccgga attgaacgag attctggata ccttcctgaa cgacgagtgc
     9181 ctcttgcatg ccatgcatat cagcacagga ctgtccatct tcgacacatc tctgttttga
     9241 aacccagctt tcttgtacaa agtggtgatg ggctgcagga attcgatatc aagcttatcg
     9301 ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     9361 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     9421 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     9481 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     9541 gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat
     9601 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     9661 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     9721 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc
     9781 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     9841 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     9901 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     9961 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    10021 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    10081 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    10141 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    10201 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt
    10261 gcggtatttc acaccgcata ggcaagtgca caaacaatac ttaaataaat actactcagt
    10321 aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt cttttacacc
    10381 atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc taagcgcatc
    10441 accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagcttt cggggctctc
    10501 ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg tcccacctgc
    10561 ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac tgagtagtat
    10621 gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga actcttggta
    10681 ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat cattgaccag
    10741 agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt tcggagtgcc
    10801 tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa ccgggtcaat
    10861 tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg aatctagagc
    10921 acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac cagaactacc
    10981 tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca cgtatactca
    11041 cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt tttcgaccga
    11101 attaattctt aatcggcaaa aaaagaaaag ctccggatca agattgtacg taaggtgaca
    11161 agctattttt caataaagaa tatcttccac tactgccatc tggcgtcata actgcaaagt
    11221 acacatatat tacgatgctg tctattaaat gcttcctata ttatatatat agtaatgtcg
    11281 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
    11341 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
    11401 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
    11461 acgcgcga
//
