LOCUS       sequence-365957-        7969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible CRE activation in zebrafish transient transgenic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7969)
  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 7969)
  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: pT2h70CRET(#325)
FEATURES             Location/Qualifiers
     source          1..7969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(50..61,62..249)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        2185..2203
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2185..2202
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          2296..2868
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2876..2895
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2923..2942
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2923..2941
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2954..3982
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             3995..4930
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             4043..4936
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     polyA_signal    4982..5103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5019..5038)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5073..5092
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(5485..5647,5648..5659)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     terminator      6017..6046
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      6068..6655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6556..6575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6667..6694)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6718..6909)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6910..7701,7702..7770))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7533..7552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7771..7873)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7874..7905)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctagaactag tggatctgct gggcttgctg aaggtagggg gtcaagaacc agaggtgtaa
       61 agtacttgag taattttact tgattactgt acttaagtat tatttttggg gatttttact
      121 ttacttgagt acaattaaaa atcaatactt ttacttttac ttaattacat ttttttagaa
      181 aaaaaagtac tttttactcc ttacaatttt atttacagtc aaaaagtact tattttttgg
      241 agatcacttc attctatttt cccttgctat taccaaacca attgaattgc gctgatgccc
      301 agtttaattt aaatgttatt tattctgcct atgaaaatcg ttttcacatt atatgaaatt
      361 ggtcagacat gttcattggt cctttggaag tgacgtcatg tcacatctat taccacaatg
      421 cacagcacct tgacctggaa attagggaaa ttataacagt caatcagtgg aagaaaatgg
      481 aggaagtatg tgattcatca gcagctgcga gcagcacagt ccaaaatcag ccacaggatc
      541 aagagcaccc gtggccgtat cttcgcagat ctgtcgactt aattaaccat ggcgtaccgg
      601 gccccccctc gactcagggg tgtcgcttgg ttatttccaa aaatcaaatt aattttatta
      661 aactattaga acgagcatgt tttgtctata tgctacagaa gataaaaaat aataggagtt
      721 aacagttata aaacaacaca ctttgtttct attgattgtt gaccacactg gggtctcatt
      781 aagttagatt aaagacacac taactgggtc aaaagcagca gattgatttc atagcaccag
      841 ggtaaacttt ctaacacttt tacggcaatc atatacatta aaattaaata cagaccacga
      901 ctgaacaagg aggatgatct ccaatattaa acaaagagac ttgtgcctat ttctctgagg
      961 gtaaacatga cctctcaagt tagcaagttg tttttaacac tacaaaaata gttaagactg
     1021 caatcccaga ataaagtatt ggttttaacc aatcaatata gtacagtaaa catccatttg
     1081 ttttgttgaa acgttaaaca aatctgacca aagctattag cttatataaa acaggtttgc
     1141 cttctatgta gctgaaaaca ccacaggccc gattttgcta ctgtgtaaaa catttcagca
     1201 agattttttt attgcatttt tttttactga atcgttcaaa cattttatca ttttagtttg
     1261 ttcattcatt gcaactggaa aaacaacaca tcacacaacc gcacatattt cagcaataag
     1321 tacaataaaa cactcaaata aaaaaaacat tttaaatctc tttgtatttt tgaccgctgt
     1381 ttcgcgtaat ttcacggtaa aactctggaa atctccacta cattcctctc agcggctcct
     1441 ctcaatgaca gctgaagaag tgacgcggct gcctgctgtg ttttgattgg tcgaattcac
     1501 tggaggcttc cagaacagtg tagagtctga acgggtgcgc gctctgctgt atttaaaggg
     1561 cgaaagagag accgcagaga aactcaaccg aagagaagcg acttgacaaa gaagaaaaga
     1621 gcagcctgac aggacttttt ccccgacgag gtgtttattc gctctattta agaatctact
     1681 gtaaggtaag tctcaatata ttgtactcta ttggctaatc agaattatat agagattata
     1741 tgtacttaat gtcaaaaaat caactttgta tatgtaatct ttttacatgt ggactgccta
     1801 tgttcatctt attttaggtc tacttgaaaa ttatatttcc cgttttcaca ataaggattt
     1861 ttaaaaaaag caatgaacag acgggcattt actttatgtt gctgacatta ttttatatga
     1921 gcataataac cataaatact agcaaatgtc ctaaatgaat ttgtgttaat gttgtctaca
     1981 aaagaaaatt agcgttttac ttgtacaact aataataact tggttattaa gagaatttca
     2041 cttgttgact agaaaaatcc tttcataatg aaacaattgc accataaatt gtataaatat
     2101 aaaattaatt ctaattgttt ttttttcctg caggaattcg atatcaagct cgccattctg
     2161 cctggggacg tcggagcaac cttgatttag gtgacactat agaatacaag ctagcttctt
     2221 gaattctttt tgcatcgatt cgaattcaag gtaggctttt gcaaaaagct ccggatcgat
     2281 cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta
     2341 aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt
     2401 cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga
     2461 caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc
     2521 ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt
     2581 ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt
     2641 tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc
     2701 tggcgtggaa atattcttat tggtagaaac aactacatcc tggtcatcat cctgcctttc
     2761 tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa
     2821 ccgggcccct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     2881 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgtaatacga ctcactatag
     2941 ggcgaattcc accatgtcca atttactgac cgtacaccaa aatttgcctg cattaccggt
     3001 cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg atcgccaggc
     3061 gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg
     3121 caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct
     3181 atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa
     3241 catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt
     3301 tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc
     3361 gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca
     3421 ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac gtatagccga
     3481 aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca
     3541 tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg
     3601 ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg atccgaataa
     3661 ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct
     3721 atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa
     3781 ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc
     3841 gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac
     3901 caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt
     3961 gcgcctgctg gaagatggcg atctcgagcc atctgctgga gacatgagag ctgccaacct
     4021 ttggccaagc ccgctcatga tcaaacgctc taagaagaac agcctggcct tgtccctgac
     4081 ggccgaccag atggtcagtg ccttgttgga tgctgagccc cccatactct attccgagta
     4141 tgatcctacc agacccttca gtgaagcttc gatgatgggc ttactgacca acctggcaga
     4201 cagggagctg gttcacatga tcaactgggc gaagagggtg ccaggctttg tggatttgac
     4261 cctccatgat caggtccacc ttctagaatg tgcctggcta gagatcctga tgattggtct
     4321 cgtctggcgc tccatggagc acccagtgaa gctactgttt gctcctaact tgctcttgga
     4381 caggaaccag ggaaaatgtg tagagggcat ggtggagatc ttcgacatgc tgctggctac
     4441 atcatctcgg ttccgcatga tgaatctgca gggagaggag tttgtgtgcc tcaaatctat
     4501 tattttgctt aattctggag tgtacacatt tctgtccagc accctgaagt ctctggaaga
     4561 gaaggaccat atccaccgag tcctggacaa gatcacagac actttgatcc acctgatggc
     4621 caaggcaggc ctgaccctgc agcagcagca ccagcggctg gcccagctcc tcctcatcct
     4681 ctcccacatc aggcacatga gtaacaaagg catggagcat ctgtacagca tgaagtgcaa
     4741 gaacgtggtg cccctctatg acctgctgct ggaggcggcg gacgcccacc gcctacatgc
     4801 gcccactagc cgtggagggg catccgtgga ggagacggac caaagccact tggccactgc
     4861 gggctctact tcatcgcatt ccttgcaaaa gtattacatc acgggggagg cagagggttt
     4921 ccctgccaca gcttgatgaa gatctgagct ccctggcgga attcggatct tattaaagca
     4981 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5041 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5101 ttatcatgtc tggtcgaggg cccatctggc ctgtgtttca gacaccaggg agtctctgct
     5161 cacgtttcct gctatttgca gcctctctat caagactaat acacctcttc ccgcatcggc
     5221 tgcctgtgag aggcttttca gcactgcagg attgcttttc agccccaaaa gagctaggct
     5281 tgacactaac aattttgaga atcagcttct actgaagtta aatctgaggt tttacaactt
     5341 tgagtagcgt gtactggcat tagattgtct gtcttatagt ttgataatta aatacaaaca
     5401 gttctaaagc aggataaaac cttgtatgca tttcatttaa tgttttttga gattaaaagc
     5461 ttaaacaaga atctctagtt ttctttcttg cttttacttt tacttcctta atactcaagt
     5521 acaattttaa tggagtactt ttttactttt actcaagtaa gattctagcc agatactttt
     5581 acttttaatt gagtaaaatt ttccctaagt acttgtactt tcacttgagt aaaatttttg
     5641 agtacttttt acacctctgt caagaactcc tggacaaacc tctgacctgt gtggaacaga
     5701 gtggatatgg gtgtctgaac agatattcac gtcttttgca gatcagaggg catttctggt
     5761 gttgtgcaat gacctgggtc caacaggtag gcgtactcgt atccaagagc attgttggac
     5821 aggaggtagt ctccattcct ataaagagga aggaatggca ccatgtagta gccgtcattg
     5881 tggccaattg gtgcattggt acgtgggtag atggaccggg gaggctgatg agttcttagc
     5941 cagcaacgaa ttcaagcttg atatcattca ggacgagcct cagactccag cgtaactgga
     6001 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     6061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     6181 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6241 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6301 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6361 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6421 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6481 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6541 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6601 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6661 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     6721 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     6781 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     6841 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     6901 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     7021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7081 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     7141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7201 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     7381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     7441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7561 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     7621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7741 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     7801 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     7861 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     7921 caggtatgat ttaaatggtc agtattgagc gatatctaga gaattcgtc
//
