LOCUS       sequence-365949-        6364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible CRE activation via mammalian cell transfection or
            mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  AUTHORS   Sinha DK, Neveu P, Gagey N, Aujard I, Le Saux T, Rampon C, Gauron C,
            Kawakami K, Leucht C, Bally-Cuif L, Volovitch M, Bensimon D, Jullien
            L, Vriz S
  TITLE     Photoactivation of the CreER T2 recombinase for conditional 
            site-specific recombination with high spatiotemporal resolution.
  JOURNAL   Zebrafish. 2010 Jun;7(2):199-204. doi: 10.1089/zeb.2009.0632.
  PUBMED    20441524
REFERENCE   2  (bases 1 to 6364)
  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"; doi: "10.1089/zeb.2009"; 
            journalName: "Zebrafish"; date: "2010-06"; volume: "7"; issue: "2"; 
            pages: "199-204"
COMMENT     Sequence Label: pCSCRET2(#323)
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             142..171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             181..210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             220..249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             259..288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             322..351
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             361..1389
                     /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             1402..2337
                     /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             1450..2343
                     /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    2416..2550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2440..2459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2494..2513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2663..2683)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2663..2681)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2702..2718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2702..2718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2715..2737)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2726..2742
                     /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)."
     promoter        complement(join(2750..2756,2757..2774,2775..2780))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2795..2816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2933..2950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3104..3692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3184..3203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3863..4654,4655..4723))
                     /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     4486..4505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4724..4828)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4854..5309)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4991..5012)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5203..5222
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5380..6364
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggagcaaaag ctcatttctg aagaggactt gaattccacc
      361 atgtccaatt tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt
      421 gatgaggttc gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat
      481 acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac
      541 cggaaatggt ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg
      601 cgcggtctgg cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt
      661 cggtccgggc tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc
      721 cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact
      781 gatttcgacc aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat
      841 ctggcatttc tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc
      901 agggttaaag atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg
      961 aaaacgctgg ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg
     1021 gtcgagcgat ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc
     1081 cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc
     1141 ctggaaggga tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt
     1201 cagagatacc tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc
     1261 cgcgctggag tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt
     1321 gtcatgaact atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa
     1381 gatggcgatc tcgagccatc tgctggagac atgagagctg ccaacctttg gccaagcccg
     1441 ctcatgatca aacgctctaa gaagaacagc ctggccttgt ccctgacggc cgaccagatg
     1501 gtcagtgcct tgttggatgc tgagcccccc atactctatt ccgagtatga tcctaccaga
     1561 cccttcagtg aagcttcgat gatgggctta ctgaccaacc tggcagacag ggagctggtt
     1621 cacatgatca actgggcgaa gagggtgcca ggctttgtgg atttgaccct ccatgatcag
     1681 gtccaccttc tagaatgtgc ctggctagag atcctgatga ttggtctcgt ctggcgctcc
     1741 atggagcacc cagtgaagct actgtttgct cctaacttgc tcttggacag gaaccaggga
     1801 aaatgtgtag agggcatggt ggagatcttc gacatgctgc tggctacatc atctcggttc
     1861 cgcatgatga atctgcaggg agaggagttt gtgtgcctca aatctattat tttgcttaat
     1921 tctggagtgt acacatttct gtccagcacc ctgaagtctc tggaagagaa ggaccatatc
     1981 caccgagtcc tggacaagat cacagacact ttgatccacc tgatggccaa ggcaggcctg
     2041 accctgcagc agcagcacca gcggctggcc cagctcctcc tcatcctctc ccacatcagg
     2101 cacatgagta acaaaggcat ggagcatctg tacagcatga agtgcaagaa cgtggtgccc
     2161 ctctatgacc tgctgctgga ggcggcggac gcccaccgcc tacatgcgcc cactagccgt
     2221 ggaggggcat ccgtggagga gacggaccaa agccacttgg ccactgcggg ctctacttca
     2281 tcgcattcct tgcaaaagta ttacatcacg ggggaggcag agggtttccc tgccacagct
     2341 tgatgaagat ctgagctccc tggcggaatt caaggcctct cgagcctcta gaactatagt
     2401 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2461 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2521 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2581 caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg
     2641 gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     2701 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2761 ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2821 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2881 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2941 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3001 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3061 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3121 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3181 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3241 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3301 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3361 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3421 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3481 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3541 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3601 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3661 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3721 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3781 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3841 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3901 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3961 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4021 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4081 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4141 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4201 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4261 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4321 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4381 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4441 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4501 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4561 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4621 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4681 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4741 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4801 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     4861 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4921 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4981 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5041 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5101 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5161 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5221 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5281 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     5341 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg
     5401 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
     5461 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
     5521 gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg
     5581 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
     5641 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
     5701 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
     5761 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
     5821 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
     5881 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
     5941 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
     6001 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
     6061 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
     6121 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
     6181 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
     6241 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
     6301 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
     6361 gaac
//
