LOCUS       sequence-401188-       11500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish transgenics; Plasmid encodes dCas9-Vp64 and could be 
            stably integrated in the zebrafish genome using Tol2 transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11500)
  AUTHORS   Pelea O, Mayes S, Ferry QRV, Fulga TA, Sauka-Spengler T
  TITLE     Specific Modulation of CRISPR Transcriptional Activators through 
            RNA-Sensing Guide RNAs in Mammalian Cells and Zebrafish Embryos
  JOURNAL   bioRxiv 2023.05.08.539738
REFERENCE   2  (bases 1 to 11500)
  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: "bioRxiv 
            2023.05.08.539738"
COMMENT     Sequence Label: Tol2_B-act_dCas9-Vp64-T2A-Citrine
FEATURES             Location/Qualifiers
     source          1..11500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             14..34
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(44..70,71..73,74..2557,2558..2563,2564..2629,
                     2630..2632,2633..4147)
                     /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             4160..4180
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4205..4354
                     /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             4382..4435
                     /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             join(4442..4444,4445..5155)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4484..4505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4745..4764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5092..5113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5188..5207)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5189..5207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5214..5348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5238..5257)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5292..5311
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5440..5577,5578..5589)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5648..5665)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5648..5664)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5657..5679)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5806..6260
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5892..5911)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6102..6123
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6286..6390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6391..6459,6460..7251)
                     /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(6609..6628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7422..8010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7911..7930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8164..8181
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8298..8319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8334..8339,8340..8357,8358..8364)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8372..8388
                     /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     8377..8399
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8396..8412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8396..8412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8431..8451
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8433..8451
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     8488..8504
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8488..8504
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(8538..8549,8550..8737)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        11452..11470
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     11452..11469
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atccgccacc atgcccaaga agaagaggaa ggtgggaagg gggatggaca agaagtactc
       61 cattgggctc gctatcggca caaacagcgt cggctgggcc gtcattacgg acgagtacaa
      121 ggtgccgagc aaaaaattca aagttctggg caataccgat cgccacagca taaagaagaa
      181 cctcattggc gccctcctgt tcgactccgg ggagacggcc gaagccacgc ggctcaaaag
      241 aacagcacgg cgcagatata cccgcagaaa gaatcggatc tgctacctgc aggagatctt
      301 tagtaatgag atggctaagg tggatgactc tttcttccat aggctggagg agtccttttt
      361 ggtggaggag gataaaaagc acgagcgcca cccaatcttt ggcaatatcg tggacgaggt
      421 ggcgtaccat gaaaagtacc caaccatata tcatctgagg aagaagcttg tagacagtac
      481 tgataaggct gacttgcggt tgatctatct cgcgctggcg catatgatca aatttcgggg
      541 acacttcctc atcgaggggg acctgaaccc agacaacagc gatgtcgaca aactctttat
      601 ccaactggtt cagacttaca atcagctttt cgaagagaac ccgatcaatg catccggagt
      661 tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc cggcggctcg aaaacctcat
      721 cgcacagctc cctggggaga agaagaacgg cctgtttggt aatcttatcg ccctgtcact
      781 cgggctgacc cccaacttta aatctaactt cgacctggcc gaagatgcca agcttcaact
      841 gagcaaagac acctacgatg atgatctcga caatctgctg gcccagatcg gcgaccagta
      901 cgcagacctt tttttggcgg caaagaacct gtcagacgcc attctgctga gtgatattct
      961 gcgagtgaac acggagatca ccaaagctcc gctgagcgct agtatgatca agcgctatga
     1021 tgagcaccac caagacttga ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa
     1081 gtacaaggaa attttcttcg atcagtctaa aaatggctac gccggataca ttgacggcgg
     1141 agcaagccag gaggaatttt acaaatttat taagcccatc ttggaaaaaa tggacggcac
     1201 cgaggagctg ctggtaaagc ttaacagaga agatctgttg cgcaaacagc gcactttcga
     1261 caatggaagc atcccccacc agattcacct gggcgaactg cacgctatcc tcaggcggca
     1321 agaggatttc tacccctttt tgaaagataa cagggaaaag attgagaaaa tcctcacatt
     1381 tcggataccc tactatgtag gccccctcgc ccggggaaat tccagattcg cgtggatgac
     1441 tcgcaaatca gaagagacca tcactccctg gaacttcgag gaagtcgtgg ataagggggc
     1501 ctctgcccag tccttcatcg aaaggatgac taactttgat aaaaatctgc ctaacgaaaa
     1561 ggtgcttcct aaacactctc tgctgtacga gtacttcaca gtttataacg agctcaccaa
     1621 ggtcaaatac gtcacagaag ggatgagaaa gccagcattc ctgtctggag agcagaagaa
     1681 agctatcgtg gacctcctct tcaagacgaa ccggaaagtt accgtgaaac agctcaaaga
     1741 ggactatttc aaaaagattg aatgtttcga ctctgttgaa atcagcggag tggaggatcg
     1801 cttcaacgca tccctgggaa cgtatcacga tctcctgaaa atcattaaag acaaggactt
     1861 cctggacaat gaggagaacg aggacattct tgaggacatt gtcctcaccc ttacgttgtt
     1921 tgaagatagg gagatgattg aagaacgctt gaaaacttac gctcatctct tcgacgacaa
     1981 agtcatgaaa cagctcaaga ggcgccgata tacaggatgg gggcggctgt caagaaaact
     2041 gatcaatggg atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga
     2101 tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga
     2161 ggacatccag aaagcacaag tttctggcca gggggacagt ctccacgagc acatcgctaa
     2221 tcttgcaggt agcccagcta tcaaaaaggg aatactgcag accgttaagg tcgtggatga
     2281 actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc gttatcgaga tggcccgaga
     2341 gaaccaaact acccagaagg gacagaagaa cagtagggaa aggatgaaga ggattgaaga
     2401 gggtataaaa gaactggggt cccaaatcct taaggaacac ccagttgaaa acacccagct
     2461 tcagaatgag aagctctacc tgtactacct gcagaacggc agggacatgt acgtggatca
     2521 ggaactggac atcaatcggc tctccgacta cgacgtggct gctatcgtgc cccagtcttt
     2581 tctcaaagat gattctattg ataataaagt gttgacaaga tccgataaag ctagagggaa
     2641 gagtgataac gtcccctcag aagaagttgt caagaaaatg aaaaattatt ggcggcagct
     2701 gctgaacgcc aaactgatca cacaacggaa gttcgataat ctgactaagg ctgaacgagg
     2761 tggcctgtct gagttggata aagccggctt catcaaaagg cagcttgttg agacacgcca
     2821 gatcaccaag cacgtggccc aaattctcga ttcacgcatg aacaccaagt acgatgaaaa
     2881 tgacaaactg attcgagagg tgaaagttat tactctgaag tctaagctgg tctcagattt
     2941 cagaaaggac tttcagtttt ataaggtgag agagatcaac aattaccacc atgcgcatga
     3001 tgcctacctg aatgcagtgg taggcactgc acttatcaaa aaatatccca agcttgaatc
     3061 tgaatttgtt tacggagact ataaagtgta cgatgttagg aaaatgatcg caaagtctga
     3121 gcaggaaata ggcaaggcca ccgctaagta cttcttttac agcaatatta tgaatttttt
     3181 caagaccgag attacactgg ccaatggaga gattcggaag cgaccactta tcgaaacaaa
     3241 cggagaaaca ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag tccggaaggt
     3301 cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg gaggcttctc
     3361 caaggaaagt atcctcccga aaaggaacag cgacaagctg atcgcacgca aaaaagattg
     3421 ggaccccaag aaatacggcg gattcgattc tcctacagtc gcttacagtg tactggttgt
     3481 ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac tgctgggcat
     3541 cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg aggcgaaagg
     3601 atataaagag gtcaaaaaag acctcatcat taagcttccc aagtactctc tctttgagct
     3661 tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag gtaacgagct
     3721 ggcactgccc tctaaatacg ttaatttctt gtatctggcc agccactatg aaaagctcaa
     3781 agggtctccc gaagataatg agcagaagca gctgttcgtg gaacaacaca aacactacct
     3841 tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg ccgacgctaa
     3901 cctcgataag gtgctttctg cttacaataa gcacagggat aagcccatca gggagcaggc
     3961 agaaaacatt atccacttgt ttactctgac caacttgggc gcgcctgcag ccttcaagta
     4021 cttcgacacc accatagaca gaaagcggta cacctctaca aaggaggtcc tggacgccac
     4081 actgattcat cagtcaatta cggggctcta tgaaacaaga atcgacctct ctcagctcgg
     4141 tggagacagc agggctgacc ccaagaagaa gaggaaggtg gaggccagcg gttccggacg
     4201 ggctgacgca ttggacgatt ttgatctgga tatgctggga agtgacgccc tcgatgattt
     4261 tgaccttgac atgcttggtt cggatgccct tgatgacttt gacctcgaca tgctcggcag
     4321 tgacgccctt gatgatttcg acctggacat gctgattaac tctagagata tcggatcagg
     4381 agagggcagg ggaagtcttc taacatgcgg ggacgtggag gaaaatcccg ggcccagatc
     4441 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4501 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4561 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4621 cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca
     4681 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4741 caaagacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4801 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4861 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4921 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4981 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5041 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5101 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     5161 cggccgcgac tctagagtat ccggactccc tatagtgagt cgtattaacc ggtgatccag
     5221 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     5281 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5341 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     5401 aggttttttg atatcccggg tttaaacgca tgcagatcta atactcaagt acaattttaa
     5461 tggagtactt ttttactttt actcaagtaa gattctagcc agatactttt acttttaatt
     5521 gagtaaaatt ttccctaagt acttgtactt tcacttgagt aaaatttttg agtacttttt
     5581 acacctctgt caagaaccat atgccggtac ccaattcgcc ctataggacg tcgtattacg
     5641 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5701 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5761 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     5821 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccaccaca cccgccgcgc
     5881 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5941 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6001 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6061 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6121 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6181 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6241 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6301 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6361 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6421 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6481 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6541 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6601 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6661 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6721 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6781 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6841 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6901 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6961 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7021 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7081 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7141 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7201 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7261 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7321 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7381 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7441 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7501 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7561 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7621 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7681 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7741 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7801 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7861 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7921 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7981 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8041 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8101 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8161 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     8221 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     8281 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     8341 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     8401 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     8461 ctggagctcc accgcggtgg cggccgctct agaactagtg gatctgctgg gcttgctgaa
     8521 ggtagggggt caagaaccag aggtgtaaag tacttgagta attttacttg attactgtac
     8581 ttaagtatta tttttgggga tttttacttt acttgagtac aattaaaaat caatactttt
     8641 acttttactt aattacattt ttttagaaaa aaaagtactt tttactcctt acaattttat
     8701 ttacagtcaa aaagtactta ttttttggag atcacttggg cccggctcga gggcctagtc
     8761 attacctata ctcaacgttt actctgagaa acttccatat tttagaggta aaatgctcag
     8821 aattgtaaaa agtcttgtca aaatacttgg aatttgccaa aaacaaaata aaagtcacat
     8881 cacctcattg tccttggtat cagcaagcac ttttaacaca atcagctgtt tttgtttata
     8941 actcccactg aaggccatgt gcattactcc caagttacac agtaaaaagt cacatgtagt
     9001 ttatcatgat cactgctgct caagcacagc tttatctgtt attcagtgtg gcaatatctg
     9061 aaatgtgtgc aaaatattat ttattgtagc tgatcaccaa cactttcagt gttttttttt
     9121 ttctactttt aatgagaatt atgttcttag gatttatcct tataaacctc tttaaaaaga
     9181 aatgttatgt ctgcatagac acgacatgaa attatgaaca gaaaatttaa ctatttaaga
     9241 tcaagacttg acatataaca tttctaatga acattaaacc tgaggtaggc ccacttgcaa
     9301 aaatgtaact gctatcactc actttataca tcattgtgta acttgtacac acaaaaatag
     9361 tgaagcaatg tggcatggaa tgcaggtccc attcattcaa agttattaaa gcactttctt
     9421 aaagtgcatt actactgagc taagaccagc tagttgctca tacaacatac tacaagtagt
     9481 gtcatcctaa tttggataaa aaaaatgtcc attttataca tggaaaagta ctcattagat
     9541 cccccattgt ttgtattacg gtatttcgtg aacacaagag gtaaatgacc tacagagctg
     9601 ctgttgtgtt agattgaaaa cacaacacag gatcatggag gcatccatca ttatgacgca
     9661 ttctgccact tgtaactctg cacagacttt ttgaaaagtt taattgagtc ttggcgttgt
     9721 cgcagaaaaa tacgaaaaat tctgtgttaa aatttgcaaa taaaaattcc agttttatag
     9781 gaaatcgcca tggccttgtt ctttaaccag ttaagcttcc ccttctttca ctctcaagtt
     9841 gcaagaagca agtgtagcaa tgtgcacgcg acagccgggt gtgtgacgct ggaccaatca
     9901 gagcacagag ctccgaaagt ttacctttta tggctagagc cgggtatgtg ccgtcatata
     9961 aaagagcgcg cccagctttt cagcctcact ttgagctcct ccacacgcag ctagtgcgga
    10021 atatcatctg cttgtaaccc attctcttaa gtcgacaacc ccccaaaccc aaggtgagtt
    10081 ggttttacag tttttaagcg tttaattata gtttctatct attaatgtta agtaaaatgt
    10141 gtaaatggat aaatatgatt tgatgaagtc tttagacagt tcataacggt ctctttgtag
    10201 ttatctacta ggtttgattt ataagtggtg gttcttttga taatctatgt tatatttaat
    10261 aaatattggc tgtatcttaa cttttttgta gcgttagtgg atgtggcagg tgagaatgtc
    10321 ggtaatttaa cgtgaccagt cgtaggcacg acattgaaca ttgaataggc cggtgtgaaa
    10381 tgggtgttaa gtcttaatat aaagttgtgt tctggctaat ctctctttaa cagccataaa
    10441 tgtgattttt aactacagtt ttgagggtgc tggtgagcat gttgcacact tgatggatag
    10501 ccggcatggg aagttctttg tgcaggcagt gctgcagcag ggtgtgacct actttagcta
    10561 gccggctaac cagctctcat ctgatgtaac gtaaacccca tcaatctaac atcggctcat
    10621 tcgtgcttta taagatcgaa gatcttgtta ttagtaacgg gtattggcat ttagttttaa
    10681 cttggttaat tttctttttg ttatgtgttc tatttaaatg tagcctgcag ccccctctgt
    10741 ctttaatgaa gctgaagatt aacgttaacg ttagtgaagt gttgtatccg ctatgaaaga
    10801 tttgaatttt agaggtgtgg tccactttaa taacggctgc ggagttctct ggctgcactt
    10861 cagccgctgt gactgtcgaa gcagagccca tcagtttttc aggatctcgg tgatttttga
    10921 ggctagacta tgaactgaac cgactgagag tgaccgtcca gaaatgcttt agtgtgaagt
    10981 ctccgctcac gaggagtccg tggactgact gcagatctgt gacgcagtaa cacccggcgg
    11041 cagacacccg ttgaaatgcg gttgtgtaat tgatcgcagg cgagggtgga agaggatgtg
    11101 aaacttcatt tgtgtaaaat ttagggagtg gtcctggtgt gatgaatgtc gaaatctgtt
    11161 cctttttact gagccctacg actctggctg agtgccacac cgccggcagc cgcaaagcgg
    11221 ctcaaaccat tgccttttat ggtaataatg agagaatgca gagggacttc ctttgtctgg
    11281 catatctgag gcgcgcgctg tcactagcgc ccaccagcgg tcagactgta gaatgcagag
    11341 caaaacagga agttgactcc agatggtcac atgcttgctg aaacgcctct ccctggcagc
    11401 agtgcaattc taacagtttc cctttctttt acagttcgcc atcctaggcc tatttaggtg
    11461 acactataga agagtacaag ctacttgttc tttttgcagg
//
