LOCUS       sequence-365701-        5813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible GFP nuclear relocation via mammalian cell 
            transfection or mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5813)
  AUTHORS   Sinha DK, Neveu P, Gagey N, Aujard I, Benbrahim-Bouzidi C, Le Saux 
            T, Rampon C, Gauron C, Goetz B, Dubruille S, Baaden M, Volovitch M, 
            Bensimon D, Vriz S, Jullien L
  TITLE     Photocontrol of protein activity in cultured cells and zebrafish 
            with one- and two-photon illumination.
  JOURNAL   Chembiochem. 2010 Mar 22;11(5):653-63. doi: 10.1002/cbic.201000008.
  PUBMED    20187057
REFERENCE   2  (bases 1 to 5813)
  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: "10.1002/cbic"; 
            journalName: "Chembiochem"; date: "2010-03-22- 22"; volume: "11"; 
            issue: "5"; pages: "653-63"
COMMENT     Sequence Label: pCSGFPnERT(#205)
FEATURES             Location/Qualifiers
     source          1..5813
                     /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"
     regulatory      131..140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(137..139,140..142,143..850)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(137..139,140..142,143..850)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(182..203)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(443..462)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     790..811
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             857..877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             899..1834
                     /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             947..1840
                     /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    1865..1999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1889..1908)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1943..1962
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2112..2132)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2112..2130)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2151..2167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2151..2167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2164..2186)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2175..2191
                     /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(2199..2205,2206..2223,2224..2229))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2244..2265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2382..2399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2633..2652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3312..4103,4104..4172))
                     /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     3935..3954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4173..4277)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4303..4758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4440..4461)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4652..4671
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4829..5813
                     /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 ttcgaattca aggcctctcg agcctctagc
      121 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      181 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      241 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      301 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      361 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      421 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      481 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      541 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      601 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      661 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      721 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
      781 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      841 cgagctgtac aggatcccca agaagaagcg caaggtggcg cccgggctcg agccatctgc
      901 tggagacatg agagctgcca acctttggcc aagcccgctc atgatcaaac gctctaagaa
      961 gaacagcctg gccttgtccc tgacggccga ccagatggtc agtgccttgt tggatgctga
     1021 gccccccata ctctattccg agtatgatcc taccagaccc ttcagtgaag cttcgatgat
     1081 gggcttactg accaacctgg cagacaggga gctggttcac atgatcaact gggcgaagag
     1141 ggtgccaggc tttgtggatt tgaccctcca tgatcaggtc caccttctag aatgtgcctg
     1201 gctagagatc ctgatgattg gtctcgtctg gcgctccatg gagcacccag tgaagctact
     1261 gtttgctcct aacttgctct tggacaggaa ccagggaaaa tgtgtagagg gcatggtgga
     1321 gatcttcgac atgctgctgg ctacatcatc tcggttccgc atgatgaatc tgcagggaga
     1381 ggagtttgtg tgcctcaaat ctattatttt gcttaattct ggagtgtaca catttctgtc
     1441 cagcaccctg aagtctctgg aagagaagga ccatatccac cgagtcctgg acaagatcac
     1501 agacactttg atccacctga tggccaaggc aggcctgacc ctgcagcagc agcaccagcg
     1561 gctggcccag ctcctcctca tcctctccca catcaggcac atgagtaaca aaggcatgga
     1621 gcatctgtac agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc tgctggaggc
     1681 ggcggacgcc caccgcctac atgcgcccac tagccgtgga ggggcatccg tggaggagac
     1741 ggaccaaagc cacttggcca ctgcgggctc tacttcatcg cattccttgc aaaagtatta
     1801 catcacgggg gaggcagagg gtttccctgc cacagcttga tgaagatctg agggcccgtt
     1861 tgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1921 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1981 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2041 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
     2101 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     2161 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     2221 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2281 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3421 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3901 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3961 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4021 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4081 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4141 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4201 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4261 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4321 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4381 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4441 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4501 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4561 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4621 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4681 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4741 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4801 cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga
     4861 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
     4921 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
     4981 accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg
     5041 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
     5101 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
     5161 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
     5221 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
     5281 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
     5341 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
     5401 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
     5461 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
     5521 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
     5581 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
     5641 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
     5701 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
     5761 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
