LOCUS       sequence-198594-       13161 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Piggybac transposon vector encoding dCAS9-VP64 and MS2-P65-HSF1 
            activator helper complex..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13161)
  AUTHORS   Li S, Zhang A, Xue H, Li D, Liu Y
  TITLE     One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of 
            Multiple Genes.
  JOURNAL   Mol Ther Nucleic Acids. 2017 Sep 15;8:64-76. doi: 
            10.1016/j.omtn.2017.06.007. Epub 2017 Jun 15.
  PUBMED    28918057
REFERENCE   2  (bases 1 to 13161)
  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: "Mol Ther 
            Nucleic Acids."; date: "2017-09-15"; volume: "8:64-76. doi"; pages: 
            " 10.1016/j.omtn.2017.06.007. Epub 2017 Jun 15"
COMMENT     Sequence Label: PB-SAM
FEATURES             Location/Qualifiers
     source          1..13161
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     4..23
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             56..103
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(104..127,128..130,131..2686,2687..2689,2690..4204)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4259..4279
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4304..4453
                     /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             4475..4528
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4529..4927
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5069..5190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5540..5574
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5698..5717)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5699..5717)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5804..5821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5975..6563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6055..6074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6734..7525,7526..7594))
                     /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     7357..7376
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7595..7699)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7767..7785
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7823..7845)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7945..7964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8045..8063
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     8045..8062
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    8129..8150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8165..8170,8171..8188,8189..8195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8203..8219
                     /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     8208..8230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8227..8243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8227..8243
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8264..8282
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   8465..8527
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        8830..10008
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9061..9999
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     9956..9976
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             10021..10407
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             10462..10482
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10498..11040
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             11065..11436
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             11452..11505
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             11509..12528
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     polyA_signal    12651..12772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        12773..13152
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagattta ccatgaaaag
       61 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat
      121 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
      181 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
      241 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
      301 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
      361 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
      421 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
      481 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
      541 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
      601 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
      661 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
      721 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
      781 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
      841 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
      901 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
      961 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     1021 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     1081 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     1141 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     1201 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     1261 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     1321 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     1381 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     1441 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     1501 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     1561 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     1621 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     1681 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     1741 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     1801 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     1861 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     1921 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     1981 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     2041 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     2101 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     2161 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     2221 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     2281 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     2341 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     2401 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     2461 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     2521 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     2581 actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct
     2641 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag
     2701 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     2761 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     2821 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     2881 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     2941 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     3001 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     3061 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     3121 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     3181 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     3241 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     3301 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     3361 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     3421 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     3481 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     3541 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     3601 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     3661 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     3721 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     3781 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     3841 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     3901 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     3961 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     4021 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     4081 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     4141 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     4201 cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc
     4261 taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat tggacgattt
     4321 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc
     4381 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga
     4441 cctggacatg ctgattaact gtacaggcag tggagagggc agaggaagtc tgctaacatg
     4501 cggtgacgtc gaggagaatc ctggcccaat ggccaagcct ttgtctcaag aagaatccac
     4561 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     4621 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     4681 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     4741 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     4801 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     4861 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     4921 gggctaaaaa tgtcgacggt accgcgggcc cgggatcctc tagatcataa tcagccatac
     4981 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     5041 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     5101 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5161 tggtttgtcc aaactcatca atgtatctta atttaaatcc atcgaagcag ctccagtcta
     5221 cacaatcgct caagacgtgt aatgctttta ttatatatta gtcacgatat ctataacaag
     5281 aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg
     5341 tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg
     5401 tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc
     5461 caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga
     5521 gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaaaa
     5581 aagatttgcg ctttactcga cctaaacttt aaacacgtca tagaatcttc gtttgacaaa
     5641 aaccacattg tggggtaccg agctcgaatt catcgatgat atcagatctg ccggtctccc
     5701 tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac
     5761 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5821 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5881 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5941 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6001 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6061 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6121 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6181 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6241 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6301 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6361 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6421 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6481 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6541 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6601 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6661 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6721 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6781 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6841 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6901 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6961 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7021 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7081 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7141 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7201 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7261 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7321 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7381 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7441 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7501 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7561 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7621 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7681 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7741 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     7801 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     7861 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     7921 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7981 tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca
     8041 tacgatttag gtgacactat agaactcgac ctcgaggctg gcacgacagg tttcccgact
     8101 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     8161 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     8221 ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa
     8281 gggaacaaaa gctggagctc gtctttgatc aaaacgcaaa tcgacgaaaa tgtgtcggac
     8341 aatatcaagt cgatgagcga aaaactaaaa aggctagaat acgacaatct cacagacagc
     8401 gttgagatat acggtattca cgacagcagg ctgaataata aaaaaattag aaactattat
     8461 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg cgtaaaattg
     8521 acgcatgtgt tttatcgatc tgtatatcga ggtttattta ttaatttgaa tagatattaa
     8581 gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt
     8641 tatttattta ttaaaaaaaa acaaaaactc aaaatttctt ctataaagta acaaaacttt
     8701 taaacattct ctcttttaca aaaataaact tattttgtac tttaaaaaca gtcatgttgt
     8761 attataaaat aagtaattag cttaacttat acataataga aacaaattat acttattaat
     8821 taatcgtgag gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     8881 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     8941 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     9001 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     9061 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
     9121 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
     9181 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
     9241 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
     9301 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     9361 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     9421 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     9481 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     9541 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     9601 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     9661 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     9721 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     9781 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     9841 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     9901 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     9961 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgaag gtttaccatg
    10021 gcttcaaact ttactcagtt cgtgctcgtg gacaatggtg ggacagggga tgtgacagtg
    10081 gctccttcta atttcgctaa tggggtggca gagtggatca gctccaactc acggagccag
    10141 gcctacaagg tgacatgcag cgtcaggcag tctagtgccc agaagagaaa gtataccatc
    10201 aaggtggagg tccccaaagt ggctacccag acagtgggcg gagtcgaact gcctgtcgcc
    10261 gcttggaggt cctacctgaa catggagctc actatcccaa ttttcgctac caattctgac
    10321 tgtgaactca tcgtgaaggc aatgcagggg ctcctcaaag acggtaatcc tatcccttcc
    10381 gccatcgccg ctaactcagg tatctacagc gctggaggag gtggaagcgg aggaggagga
    10441 agcggaggag gaggtagcgg acctaagaaa aagaggaagg tggcggccgc tggatcccct
    10501 tcagggcaga tcagcaacca ggccctggct ctggccccta gctccgctcc agtgctggcc
    10561 cagactatgg tgccctctag tgctatggtg cctctggccc agccacctgc tccagcccct
    10621 gtgctgaccc caggaccacc ccagtcactg agcgctccag tgcccaagtc tacacaggcc
    10681 ggcgagggga ctctgagtga agctctgctg cacctgcagt tcgacgctga tgaggacctg
    10741 ggagctctgc tggggaacag caccgatccc ggagtgttca cagatctggc ctccgtggac
    10801 aactctgagt ttcagcagct gctgaatcag ggcgtgtcca tgtctcatag tacagccgaa
    10861 ccaatgctga tggagtaccc cgaagccatt acccggctgg tgaccggcag ccagcggccc
    10921 cccgaccccg ctccaactcc cctgggaacc agcggcctgc ctaatgggct gtccggagat
    10981 gaagacttct caagcatcgc tgatatggac tttagtgccc tgctgtcaca gatttcctct
    11041 agtgggcagg gaggaggtgg aagcggcttc agcgtggaca ccagtgccct gctggacctg
    11101 ttcagcccct cggtgaccgt gcccgacatg agcctgcctg accttgacag cagcctggcc
    11161 agtatccaag agctcctgtc tccccaggag ccccccaggc ctcccgaggc agagaacagc
    11221 agcccggatt cagggaagca gctggtgcac tacacagcgc agccgctgtt cctgctggac
    11281 cccggctccg tggacaccgg gagcaacgac ctgccggtgc tgtttgagct gggagagggc
    11341 tcctacttct ccgaagggga cggcttcgcc gaggacccca ccatctccct gctgacaggc
    11401 tcggagcctc ccaaagccaa ggaccccact gtctcctgta caggcagtgg agagggcaga
    11461 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaaccat gaaaaagcct
    11521 gaactcaccg ctacctctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
    11581 ctgatgcagc tctccgaggg cgaagaatct cgggctttca gcttcgatgt gggagggcgt
    11641 ggatatgtcc tgcgggtgaa tagctgcgcc gatggtttct acaaagatcg ctatgtttat
    11701 cggcactttg catccgccgc tctccctatt cccgaagtgc ttgacattgg ggagttcagc
    11761 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcaccttgca agacctgcct
    11821 gaaaccgaac tgcccgctgt tctccagccc gtcgccgagg ccatggatgc catcgctgcc
    11881 gccgatctta gccagaccag cgggttcggc ccattcggac ctcaaggaat cggtcaatac
    11941 actacatggc gcgatttcat ctgcgctatt gctgatcccc atgtgtatca ctggcaaact
    12001 gtgatggacg acaccgtcag tgcctccgtc gcccaggctc tcgatgagct gatgctttgg
    12061 gccgaggact gccccgaagt ccggcacctc gtgcacgccg atttcggctc caacaatgtc
    12121 ctgaccgaca atggccgcat aacagccgtc attgactgga gcgaggccat gttcggggat
    12181 tcccaatacg aggtcgccaa catcttcttc tggaggccct ggttggcttg tatggagcag
    12241 cagacccgct acttcgagcg gaggcatccc gagcttgcag gatctcctcg gctccgggct
    12301 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
    12361 gatgcagctt gggctcaggg tcgctgcgac gcaatcgtcc ggtccggagc cgggactgtc
    12421 gggcgtacac aaatcgcccg cagaagcgct gccgtctgga ccgatggctg tgtggaagtg
    12481 ctcgccgata gtggaaacag acgccccagc actcgtccta gggcaaagga tctgcagtaa
    12541 aaacgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
    12601 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgtcgttgtt aacttgttta
    12661 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    12721 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tagacattga
    12781 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
    12841 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
    12901 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
    12961 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
    13021 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
    13081 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
    13141 gctattacca tggtcgaggt g
//
