LOCUS       sequence-363955-        9627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible GAL4 activation in zebrafish permanent 
            transgenic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9627)
  AUTHORS   Feng Z, Nam S, Hamouri F, Aujard I, Ducos B, Vriz S, Volovitch M, 
            Jullien L, Lin S, Weiss S, Bensimon D
  TITLE     Optical Control of Tumor Induction in the Zebrafish.
  JOURNAL   Sci Rep. 2017 Aug 23;7(1):9195. doi: 10.1038/s41598-017-09697-x.
  PUBMED    28835665
REFERENCE   2  (bases 1 to 9627)
  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: "Sci Rep."; 
            date: "2017-08-23"; volume: "7(1):9195. doi"; pages: " 
            10.1038/s41598-017-09697-x"
COMMENT     Sequence Label: pT22gCfpUbiG4FER4P2A(#249)
FEATURES             Location/Qualifiers
     source          1..9627
                     /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    397..518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(538..1245,1246..1248,1249..1251))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(577..598)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     926..945
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1185..1206
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2012..2023)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5657..6100
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MGKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKT
                     KRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVN
                     KDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             6188..6208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7175..7231
                     /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"
     polyA_signal    7273..7394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(7490..7627,7628..7639)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7707..7724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7878..8466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7958..7977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8637..9428,9429..9497))
                     /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     9260..9279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9498..9602)
                     /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 tcaggaccag acatgataag atacattgat gagtttggac
      421 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      481 ctttatttgt aaccattata agctgcaata aacaagttgc gtgcgcattt atttaaggta
      541 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
      601 atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc
      661 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg
      721 cacgctgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc cgttcttctg
      781 cttgtcggcg gtgatataga cgttgtggct gatgtagttg tactccagct tgtgccccag
      841 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
      901 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
      961 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     1021 gtcggggtag cggctgaagc actgcacgcc ccaggtcagg gtggtcacga gggtgggcca
     1081 gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc
     1141 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     1201 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tgtcggctgg
     1261 cgaccggtgg atcatccccc gcaagtctga ctgagtggaa gtttcagttc aatggaaggg
     1321 agaaaagagc cagcctttat atatgtcatg tattgctgct gtctgcatgc gatgaaccta
     1381 gagcttactt tttctgtcaa gtaaggcaaa agcagagaac ctttgtcttg cattgtgttt
     1441 gatgttgtgc tgtgacagtc atttcttagt tcatgctggt acttttgttc catagcttta
     1501 cccaaagagt tatccagcat tcccagttat tccaaaaact caatgtcaca aattttccag
     1561 tccattgctt agcagcgttg accccggaga ccctgatgtg gtaatttaca gtgcaattca
     1621 gttgctggtc caaacacttt gtaacagttt aaagaattgt ggttactagt aaatccttaa
     1681 tttggcatca tctaggtgtc attctttgcc acttttgaat cacacaaatg aaccaaacaa
     1741 tggttagcta gcatcaaaca tttaccaaat aatctcatca gtgtcacccc aatgtttaag
     1801 aagccacatt ttatgtcccc agatatatta atattatttt atatataatt taaaatagta
     1861 atgtggtgtt actgtatttg tcatataacg caatatatac agtatgtctt tatctgtatc
     1921 aattttgtgg tcacaccatc aataaactcc agtttaatta tttaaaattg ggtagtgata
     1981 accatataaa ggaacagtgg ctagttccat gttgtagctc ccaacctgct ttcaggtgat
     2041 ccatgcagtg tcgtataaaa aggtaacaat ttttaacctt ggaaagcaag tacacttaag
     2101 gtcccagatc tgaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta
     2161 ggtcgagacc agcaaagttc tagaatttgt cgaaacattt atgttatata tttcctgaaa
     2221 aaaattctga gtaagttctt aagtgttatt gccagcaaca taaacaacag acggcaaaat
     2281 gaataaatga taacaaagca gtaggcttaa ataaacctaa tttttatagg ctgttctcta
     2341 caaccctcaa acagtgatta gttttgtact tataaacttg ccctttcatt catatttcaa
     2401 gaaaattggt tcagaagatc tggatattct agcagttgtt caagctcatg gagggatcag
     2461 tgacctgatt ccacaatgac taggcctaat ccagaaatta gatgactgtc aacataaaaa
     2521 ggcacagcac tcactagctg ccctatatat tttattatat tttacatata ttattttatt
     2581 tatttagctc tgagtgctgt actttctggt taaagaaaac tgcttacaac agctaacctg
     2641 tactacctca ggctcaggga atttggaaca ggtttgtctg gtttgtttct ttaaccatgc
     2701 atgcttgttt tcaactatgg caacacagtc acatgggaca ttacagaaat gatttgtcga
     2761 tgacatgcga cttttcttta ataaagcgca aagatcccaa aaagcaaact tttaacaaaa
     2821 atcatataat tatattttca atccagcttt gtagcaactt tgtgctgctg ttcactcagc
     2881 aacagatagt cagtataagg tcagtgtgtc tcaaagcagt gccatctgtt tcacacattg
     2941 cgttctatat ataagtgtgc tggttgacac gacactgtat aaggcctagg ctaaaacaca
     3001 aacaatgtag aatgacactg tgtttttttt gtaaacaaat gttgtttttg gttaaacatc
     3061 tttgtgaaaa catcctcctg tcatgtattt gctatattca aatgttaaac ccgtgcagaa
     3121 tagaacatat acaaaaaaaa acaacacaac acatttttaa acattattaa atatcaagta
     3181 ttgctggcag ttctgtttct gttttacagt accctttgcc acagttctcc gcttttcctg
     3241 gtccagattc cacaagtctg attcaccaat agcaaagcga ataaacaacc aaagcagcca
     3301 atcactgctt gtagactgtc ctgcgagacc ggcccattcc agcacattct ggaaacttcc
     3361 tttatatgat aattataaat acatttaaat tattgataca aaacatgtaa ttcctagaac
     3421 ataaccatag caatcattag ttttcagggt aattatgtat ttttaggatt tgactgcgga
     3481 aagatctggt catgtgacgt ctcatgaacg tcacggccct gggtttctat aaatacagta
     3541 ggactctcga ccatcggcag atttttcgaa gaagaagatc agtttcagga gccgtactgt
     3601 tccgtttcaa cgcaaatatt aacggtaaga gcgaatttcc tagtttgttt tcatgccatt
     3661 ctttaaaacc atagcgtatt actttaatta tagtaaactt tcgctttctt tattacaaga
     3721 gacgttttgt gttgattctc cgcggacatt ttcggtcaga caatcagaaa atgaccgcgg
     3781 aggaccagta acttgcatta cacgtaagtt aaatcttcgt gtattaaaat ggttaggttg
     3841 ttaacgtcaa ataggttacc gtgtttgcgt gtgatcaggt tggttttgtt agatttttgt
     3901 cagtattttt aatttatttg ttttagttta tttatttttt ttgctgaatc atagtttgtg
     3961 aacaaagaac ccggatgtta catacagtac agccgccatg ttacagagag ttataactta
     4021 atcattttaa aaataatttt gccttacttt tagtttgtca tgttgagaaa tgaggaaatg
     4081 ttaaaatgag gaaatatcca attaatttaa tatatcaaaa taatccatga ttacaatgca
     4141 ctgaactgga gaaaattaag atgttttcta gtgtcatgaa acaaatgtaa gagatgtaca
     4201 ttgtagatgt tttatgtcaa gaattggcta gttgatgcag catactggcg atactcagtt
     4261 gtaataacag taacgttaca tgttaataga ctactgagta tgctgttctg tctatgtatg
     4321 ctctgtaagc tacgagaagg acttttttaa acagtaaagg gtgcaatatt tttacaaatt
     4381 gaattaaata aaggctgtct attaagtaat atgcttgata tttttcttac ttgatcggaa
     4441 ataagaaaaa atataaacgt tgttgctcta aaaatcctag ttcagtttag ccaaccacaa
     4501 ataccttttt gttcctccaa cagttttttt ttcttctcta taatatttgg cagtctatag
     4561 tactccaaat gtttccccac agtctaacta attggtacag ccaaaatcat gacacttatt
     4621 gcaataataa ttttggttca ttggcattgt tgatagcctg tgccactaat atggtcgatt
     4681 gatcatgctt caggaagaaa actatattgt ttgatgtaag attattaaat cttcacctgc
     4741 ttccattaca aactattccc atcttattga attctggtat gtcttaaagg attagttcac
     4801 ttcccaaatc aaaatttact tagccttttt tcatccatga tccctttttt tcatcattaa
     4861 tgaagaaatt gtttttgaaa aagtttcaag atttttttct ctatattgtg gagcttgtta
     4921 gtttaaaatt ccaaaatgca atatgtggct tcaaatggtt ctaaatgatc ccagtcaagg
     4981 aataacagtc ttatctaatg aaaccattag acctttaaaa ataaaaaaaa taaaagtatt
     5041 tatttttaaa tgactgagtg attaagttga atttcagcgt ttccttactg tgtagaagtc
     5101 cttccttact ggccccaccc tttggttctc tgccaatctg ctacctaatg taatgttgtg
     5161 gaacattatt attctttatt tcttaatttt ttatttttta ttttaaaaca atgtaaactg
     5221 cacagatgtg cagtttgttt aaaatggcca atgctttgga aatgcatgac ataattagat
     5281 ttcatgatgc acaaagccaa atctcagagc ttgtgcaaaa tgagctatca tttcactagg
     5341 taagacccta aattttcata taggatcatt tggacaattt tgctgcaggt aaaatgcatt
     5401 ctatagtcca ctgtcagcca ttgttttgga tagtatttat ttttctctac aagtatagtc
     5461 aatagttttc tattatttta aaggtttgta acatttaagg gtgaccaaat gcaaagtaaa
     5521 atttcatttt cgggtgaact atctcgttta acatgggaga agtgcaaaac atacattatt
     5581 ggctagaaca ttgtagtatt ttttaaatgg aaatgtgtga ttgctaatct tactttgaat
     5641 ttgtttacag ggatccatgg ggaagctact gtcttctatc gaacaagcct gcgatatttg
     5701 ccgacttaaa aagctgaagt gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa
     5761 caactgggag tgtcgctact ctcccaaaac caaaaggtct ccgctgacta gggcacatct
     5821 gacagaagtg gaatcaaggc tagaaagact ggaacagcta tttctactga tttttccccg
     5881 cgaagacctt gacatgattt tgaaaatgga ttctttacag gatataaaag cactgctaac
     5941 aggattattc gtacaagata atgtgaataa agatgccgtc acagatagat tggcttcagt
     6001 ggagactgat atgcctctaa cattgagaca gcatagaata agtgcgacat catcatcgga
     6061 ggaaagtagt aacaaaggtc aaagacagct cactgtatcg cccgccgacg cactggacga
     6121 cttcgacctg gacatgctgc ctgctgatgc tctcgatgac tttgatctcg atatgctgcc
     6181 cgggccagag aacctgtact tccaggggct cgagccctct gccggagaca tgagagctgc
     6241 aaacctttgg ccgagcccgc tgatgattaa acgctctaag aagaacagcc tggccttgtc
     6301 cctgaccgcc gaccagatgg tcagtgcctt gttggatgcc gagcccccca tactctattc
     6361 cgagtatgat cctaccagac ccttcagtga ggcttcgatg atgggcttac tgaccaacct
     6421 ggcagacagg gagctggttc acatgattaa ctgggccaag agggtgccag gctttgtgga
     6481 tttgaccctc cacgatcagg tccacctcct agaatgtgcc tggctagaga tcctgatgat
     6541 tggtctcgtc tggcgcagca tggagcaccc agtgaagcta ctgtttgctc ctaacttgct
     6601 cttggacagg aaccagggaa aatgtgtaga gggcatggtg gagattttcg acatgctgct
     6661 ggctacatca tctcggttcc gcatgatgaa tctgcaggga gaggaatttg tgtgcctcaa
     6721 atctattatt ttgcttaatt ctggagttta cacatttctg tccagcaccc tgaagtctct
     6781 ggaggaaaag gaccatatcc atcgagtgct ggacaagatc acagacactt tgattcacct
     6841 gatggctaag gctggcctga ccctgcagca gcagcatcag agactcgcac agcttctcct
     6901 gatcctctcc cacatcaggc acatgagtaa caaaggcatg gagcatctgt actctatgaa
     6961 gtgcaagaac gtggtgcccc tctatgatct gctgctggag gcggcggacg ctcaccgcct
     7021 acatgcgccc actagccgtg gaggggcatc cgtggaggaa acagaccaaa gccacttggc
     7081 tactgcgggc tctacttcat ctcattcctt gcaaaagtat tacatcacgg gggaggcaga
     7141 gggtttccct gccacagcac caggttccgg cagcgctaca aacttctctc tgctcaagca
     7201 ggctggagat gtggaagaaa accctgggcc cagctacgta agatctgaga tctgatcatt
     7261 taaatgacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     7321 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc atattaagct
     7381 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     7441 tgtgggaggt tttttcctgc aggactctag tcgcggccgc tcttccgcta atactcaagt
     7501 acaattttaa tggagtactt ttttactttt actcaagtaa gattctagcc agatactttt
     7561 acttttaatt gagtaaaatt ttccctaagt acttgtactt tcacttgagt aaaatttttg
     7621 agtacttttt acacctctgc cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7681 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7801 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7861 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     8041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     8101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8281 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     8341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8761 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     9001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     9061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9601 cgcacatttc cccgaaaagt gccacct
//
