LOCUS       sequence-370311-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible mCherry 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 5765)
  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 5765)
  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: pCSmChnERT(#99)
FEATURES             Location/Qualifiers
     source          1..5765
                     /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      86..95
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             92..802
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELLYK"
     primer_bind     complement(237..255)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     505..524
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             809..829
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             851..1786
                     /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             899..1792
                     /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    1817..1951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1841..1860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1895..1914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2064..2084)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2064..2082)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2103..2119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2103..2119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2116..2138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2127..2143
                     /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(2151..2157,2158..2175,2176..2181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2196..2217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2334..2351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2505..3093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2585..2604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3264..4055,4056..4124))
                     /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     3887..3906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4125..4229)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4255..4710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4392..4413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4604..4623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4781..5765
                     /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 atcccgccac catggtgagc aagggcgagg aggataacat
      121 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      181 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
      241 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
      301 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
      361 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
      421 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
      481 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
      541 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
      601 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
      661 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
      721 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
      781 catggacgag ctgctgtaca aatcgatccc caagaagaag cgcaaggtgg cgcccgggct
      841 cgagccatct gctggagaca tgagagctgc caacctttgg ccaagcccgc tcatgatcaa
      901 acgctctaag aagaacagcc tggccttgtc cctgacggcc gaccagatgg tcagtgcctt
      961 gttggatgct gagcccccca tactctattc cgagtatgat cctaccagac ccttcagtga
     1021 agcttcgatg atgggcttac tgaccaacct ggcagacagg gagctggttc acatgatcaa
     1081 ctgggcgaag agggtgccag gctttgtgga tttgaccctc catgatcagg tccaccttct
     1141 agaatgtgcc tggctagaga tcctgatgat tggtctcgtc tggcgctcca tggagcaccc
     1201 agtgaagcta ctgtttgctc ctaacttgct cttggacagg aaccagggaa aatgtgtaga
     1261 gggcatggtg gagatcttcg acatgctgct ggctacatca tctcggttcc gcatgatgaa
     1321 tctgcaggga gaggagtttg tgtgcctcaa atctattatt ttgcttaatt ctggagtgta
     1381 cacatttctg tccagcaccc tgaagtctct ggaagagaag gaccatatcc accgagtcct
     1441 ggacaagatc acagacactt tgatccacct gatggccaag gcaggcctga ccctgcagca
     1501 gcagcaccag cggctggccc agctcctcct catcctctcc cacatcaggc acatgagtaa
     1561 caaaggcatg gagcatctgt acagcatgaa gtgcaagaac gtggtgcccc tctatgacct
     1621 gctgctggag gcggcggacg cccaccgcct acatgcgccc actagccgtg gaggggcatc
     1681 cgtggaggag acggaccaaa gccacttggc cactgcgggc tctacttcat cgcattcctt
     1741 gcaaaagtat tacatcacgg gggaggcaga gggtttccct gccacagctt gatgaagatc
     1801 tgagggcccg tttgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     1861 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1921 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1981 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg
     2041 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     2101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2161 gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2701 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2941 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     4261 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4321 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4381 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4441 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4501 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4561 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4621 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4681 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     4741 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat
     4801 ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat
     4861 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat
     4921 ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag
     4981 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc
     5041 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata
     5101 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat
     5161 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc
     5221 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac
     5281 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg
     5341 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg
     5401 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat
     5461 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact
     5521 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg
     5581 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
     5641 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
     5701 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
     5761 ggaac
//
