LOCUS       sequence-363989-        8304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible apoptosis activation and imaging (tEosFP) in 
            zebrafish permanent transgenic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8304)
  AUTHORS   Feng Z, Zhang W, Xu J, Gauron C, Ducos B, Vriz S, Volovitch M, 
            Jullien L, Weiss S, Bensimon D
  TITLE     Optical control and study of biological processes at the single-cell
            level in a live organism.
  JOURNAL   Rep Prog Phys. 2013 Jul;76(7):072601. doi: 
            10.1088/0034-4885/76/7/072601. Epub 2013 Jun 14.
  PUBMED    23764902
REFERENCE   2  (bases 1 to 8304)
  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: "Rep Prog 
            Phys."; date: "2013-07"; volume: "76(7):072601. doi"; pages: " 
            10.1088/0034-4885/76/7/072601. Epub 2013 Jun 14"
COMMENT     Sequence Label: pT22iuasvApoWtEos3N(#486)
FEATURES             Location/Qualifiers
     source          1..8304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(150..170)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(150..168)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    join(179..190,191..378)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    429..550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(570..1277,1278..1280,1281..1283))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(609..630)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     958..977
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1217..1238
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2044..2055)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2397..2438
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             2907..2927
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2928..2948
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2952..3632
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MVSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEV
                     KEGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIA
                     RNDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDITMALL
                     LEGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     CDS             3660..3716
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5100..5120
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6093..6214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(6334..6471,6472..6483)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     rep_origin      complement(6555..7143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6635..6654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7314..8105,8106..8174))
                     /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     7937..7956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8175..8279)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccc gatactccag cttttgttcc ctttagtgag ggttaattgc gcgttacaca
      181 gaggtgtaaa gtacttgagt aattttactt gattactgta cttaagtatt atttttgggg
      241 atttttactt tacttgagta caattaaaaa tcaatacttt tacttttact taattacatt
      301 tttttagaaa aaaaagtact ttttactcct tacaatttta tttacagtca aaaagtactt
      361 attttttgga gatcacttcc tcatagggat aacagggtaa tggccggcct tttcaggacc
      421 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      481 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      541 taaacaagtt gcgtgcgcat ttatttaagg tacagctcgt ccatgccgag agtgatcccg
      601 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
      661 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
      721 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
      781 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg
      841 ctgatgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      901 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      961 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     1021 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     1081 ccccaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     1141 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     1201 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     1261 agctcctcgc ccttgctcac catgtcggct ggcgaccggt ggatcatccc ccgcaagtct
     1321 gactgagtgg aagtttcagt tcaatggaag ggagaaaaga gccagccttt atatatgtca
     1381 tgtattgctg ctgtctgcat gcgatgaacc tagagcttac tttttctgtc aagtaaggca
     1441 aaagcagaga acctttgtct tgcattgtgt ttgatgttgt gctgtgacag tcatttctta
     1501 gttcatgctg gtacttttgt tccatagctt tacccaaaga gttatccagc attcccagtt
     1561 attccaaaaa ctcaatgtca caaattttcc agtccattgc ttagcagcgt tgaccccgga
     1621 gaccctgatg tggtaattta cagtgcaatt cagttgctgg tccaaacact ttgtaacagt
     1681 ttaaagaatt gtggttacta gtaaatcctt aatttggcat catctaggtg tcattctttg
     1741 ccacttttga atcacacaaa tgaaccaaac aatggttagc tagcatcaaa catttaccaa
     1801 ataatctcat cagtgtcacc ccaatgttta agaagccaca ttttatgtcc ccagatatat
     1861 taatattatt ttatatataa tttaaaatag taatgtggtg ttactgtatt tgtcatataa
     1921 cgcaatatat acagtatgtc tttatctgta tcaattttgt ggtcacacca tcaataaact
     1981 ccagtttaat tatttaaaat tgggtagtga taaccatata aaggaacagt ggctagttcc
     2041 atgttgtagc tcccaacctg ctttcaggtg atccatgcag tgtcgtataa aaaggtaaca
     2101 atttttaacc ttggaaagca agtacactta aggtcccttt gctggtctcg acctagtgaa
     2161 gagcattttc tgtgtgtaga atagtgtgtt aggggtagct tctagactgg acctcggtgg
     2221 cttctaatcc gtgagtccta gcgggtgaca gccctccgct ttcacaggcg gaggagagtc
     2281 ctccgtaggg ttcctcggag tactgtcctc cggctagtgc gaccgtctat ataagcagag
     2341 ctggtttagt gaaccgtcag atccggaggc tgccaaccag ctccagcgga tccatgatgg
     2401 ggagtagcaa gagcaagcct aaagacccta gccagagaag caggggatct accggtgcat
     2461 ctgttgtagc tgatcttccg tcttatggac ctatcgcttt cgatcaggtg gatggcgtca
     2521 cgaacgcgac ccagctattc aaaaataata taaacaaaac cagaatgaac gatttaaacc
     2581 gcgaggagac acgattaaag accttcaccg actggccgct agattggctg gataaacgcc
     2641 agctggccca aaccggcatg tacttcacac acgctggcga caaagttaaa tgctttttct
     2701 gcggcgtgga aatcggttgc tgggagcagg aggatcagcc cgtgccggaa catcagcgat
     2761 ggtcgcccaa ctgtccactg ttgcgcagga gaactaccaa caatgtgccg atcaatgccg
     2821 aagcattaga tcgcatcctg ccgccaataa gctacgacat ctgcggcgcc aacgactcga
     2881 cgctagagat gaggagatcc catcgcccta aaaagaagag aaaagtgcca aagaagaaac
     2941 gcaaggtcgc catggtgagt gcgattaagc cagacatgaa gatcaacctc cgtatggaag
     3001 gcaacgtaaa cgggcaccat tttgtgatcg acggagatgg tacaggcaag ccttttgagg
     3061 gaaaacagag tatggatctt gaagtcaaag agggcggacc tctgcctttt gcctttgaca
     3121 tcctgactac cgctttccat tacggcaaca gggtattcgc cgaatatcca gaccacatac
     3181 aagactattt taagcagtcg tttcctaaag gatattcgtg ggaacgcagc ttgactttcg
     3241 aggacggagg catttgtatc gccagaaacg acataacaat ggaaggggac actttctata
     3301 ataaagttcg atttcacggt gtaaactttc ccgccaatgg tccagttatg cagaagaaaa
     3361 cgctgaaatg ggagccctcc actgagaaaa tgtatgtgcg tgatggagtg ctgacgggtg
     3421 atattacaat ggctttgttg cttgaaggaa atgcccatta ccgatgtgac ttcagaacta
     3481 cttacaaagc taaagagaag ggtgtcaagt taccaggcta ccactttgtt gaccactgta
     3541 ttgagatttt aagccatgac aaagattaca acaaggttaa gctgtatgag cacgctgttg
     3601 ctcattctgg attgcctgac aatgccagga gatctcccaa gaagaggaag gtgagatctg
     3661 ccaccaattt tagcctcctg aaacaagcag gcgacgttga ggagaatcca ggaccgtctc
     3721 ccatgatgga cgaggcggac cgccagctcc tgagaagatg cagggtgcgc ctagtgagcg
     3781 agctgcaagt cgcggagctg tgggacgctc tgctgagtcg agagctgttc acgcgcgaca
     3841 tgatcgagga tattcagcag gcaggatctg gctctcggcg ggatcaggct aggcagctcg
     3901 tcacagacct ggagactaga ggccgccagg ctctgcctct gttcatctcc tgcttagagg
     3961 acacaggcca aggcaccctg gcttcactct tgcaaagcgg tcggcaagca gccaagcaag
     4021 acccagaggc tgttaaaccc ctagaccatc tggtgcctgt ggtcctggga ccaatgggac
     4081 tcacagcaaa ggagcagaga gtagtgaagc tggacccgtc acagcctgcc gtgggaaacc
     4141 tgaccccagt ggtgctcgga cctgaagaac tctggcctgc tcgcctgaag ccagaggttc
     4201 tcagaccaga aacacctaga cccgtggaca ttggttctgg tggagcccat gatgtctgtg
     4261 ttccagggaa gatcagggga catgcagata tggcatacac cctggactct gatccctgcg
     4321 gccactgtct catcatcaac aatgtgaact tctgcccttc ctcggggctc ggcacacgca
     4381 cgggctccaa cttggaccgt gacaaactgg agcaccgatt ccgctggctg cgcttcatgg
     4441 tggaggtgaa gaacgacctg actgctaaga aaatggtcac ggctttgatg gagatggcac
     4501 accggaacca ccgtgccctg gactgctttg tggtggtcat cctctctcat ggctgccagg
     4561 ccagccacct ccagttccca ggtgctgttt atgggacaga tggatgctcc gtgtccattg
     4621 agaaaattgt gaacatcttt aacggaagcg gctgccctag cctgggaggg aagcccaaac
     4681 tgttcttcat ccaggcttgc ggaggtgagc agaaagacca cggctttgag gtggcttgca
     4741 cttcctctca gggcagaacc ttggacagtg actctgagcc tgatgctgtc ccctatcagg
     4801 aaggcccaag acccttggac caactggatg ctgtgtcaag tttgcctacc cctagcgaca
     4861 tccttgtgtc ctactccacc ttcccaggat ttgtgtcttg gagggacaag aaaagtggct
     4921 cctggtacat cgagactttg gacggtattc tggaacagtg ggctcgctct gaggatctgc
     4981 agtccctcct tctcagggtt gccaatgctg tttctgcgaa agggacttac aagcagattc
     5041 ctggctgttt taacttcctg cggaaaaagc tgtttttcaa aacttctgtg cccgggccag
     5101 agaacctgta cttccagggg ctcgagccct ctgccggaga catgagagct gcaaaccttt
     5161 ggccgagccc gctgatgatt aaacgctcta agaagaacag cctggccttg tccctgaccg
     5221 ccgaccagat ggtcagtgcc ttgttggatg ccgagccccc catactctat tccgagtatg
     5281 atcctaccag acccttcagt gaggcttcga tgatgggctt actgaccaac ctggcagaca
     5341 gggagctggt tcacatgatt aactgggcca agagggtgcc aggctttgtg gatttgaccc
     5401 tccacgatca ggtccacctc ctagaatgtg cctggctaga gatcctgatg attggtctcg
     5461 tctggcgcag catggagcac ccagtgaagc tactgtttgc tcctaacttg ctcttggaca
     5521 ggaaccaggg aaaatgtgta gagggcatgg tggagatttt cgacatgctg ctggctacat
     5581 catctcggtt ccgcatgatg aatctgcagg gagaggaatt tgtgtgcctc aaatctatta
     5641 ttttgcttaa ttctggagtt tacacatttc tgtccagcac cctgaagtct ctggaggaaa
     5701 aggaccatat ccatcgagtg ctggacaaga tcacagacac tttgattcac ctgatggcta
     5761 aggctggcct gaccctgcag cagcagcatc agagactcgc acagcttctc ctgatcctct
     5821 cccacatcag gcacatgagt aacaaaggca tggagcatct gtactctatg aagtgcaaga
     5881 acgtggtgcc cctctatgat ctgctgctgg aggcggcgga cgctcaccgc ctacatgcgc
     5941 ccactagccg tggaggggca tccgtggagg aaacagacca aagccacttg gctactgcgg
     6001 gctctacttc atctcattcc ttgcaaaagt attacatcac gggggaggca gagggtttcc
     6061 ctgccacagc accaggatct acgtagacat gataagatac attgatgagt ttggacaaac
     6121 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     6181 atttgtaacc atattaagct gcaataaaca agttaacaac aacaattgca ttcattttat
     6241 gtttcaggtt cagggggagg tgtgggaggt tttttcctgc aggactctag tcgcggccgc
     6301 tagggataac agggtaatgg ccgctcttcc gctaatactc aagtacaatt ttaatggagt
     6361 acttttttac ttttactcaa gtaagattct agccagatac ttttactttt aattgagtaa
     6421 aattttccct aagtacttgt actttcactt gagtaaaatt tttgagtact ttttacacct
     6481 ctgccctgca tcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6541 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6601 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6721 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6961 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7321 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7381 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7441 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7501 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7561 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7621 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7681 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7741 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7801 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7861 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7921 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7981 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8041 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8101 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8161 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8221 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8281 acatttcccc gaaaagtgcc acct
//
