LOCUS       sequence-365948-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible Kaede 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 5752)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 5752)
  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"
COMMENT     Sequence Label: pCSKaedenERT(#100)
FEATURES             Location/Qualifiers
     source          1..5752
                     /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             796..816
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             838..1773
                     /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             886..1779
                     /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    1804..1938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1828..1847)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1882..1901
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2051..2071)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2051..2069)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2090..2106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2090..2106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2103..2125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2114..2130
                     /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(2138..2144,2145..2162,2163..2168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2183..2204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2321..2338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2492..3080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2572..2591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3251..4042,4043..4111))
                     /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     3874..3893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4112..4216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4242..4697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4591..4610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4768..5752
                     /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 atccaccatg agtctgatta aaccagaaat gaagatcaag
      121 ctgcttatgg aaggcaatgt aaacgggcac cagtttgtta ttgagggaga tggaaaaggc
      181 catccttttg agggaaaaca gagtatggac cttgtagtca aagaaggcgc acctctccct
      241 tttgcctacg atatcttgac aacagcattc cattatggta acagggtttt tgctaaatac
      301 ccagaccata taccagacta cttcaagcag tcgtttccca aagggttttc ttgggagcga
      361 agcctgatgt tcgaggacgg gggcgtttgc atcgctacaa atgacataac actgaaagga
      421 gacacttttt ttaacaaagt tcgatttgat ggcgtaaact ttcccccaaa tggtcctgtt
      481 atgcagaaga agactctgaa atgggaggca tccactgaga aaatgtacct gcgagatggc
      541 gttctgacag gagacatcac gatggcactc ttgctgaaag gcgatgtgca ctatcggtgt
      601 gactttcgca ctacctacaa gtccaggcag gaaggagtca aacttcctgg gtatcacttc
      661 gtggatcact gcatcagcat tctgagacac gacaaggact acaacgaggt gaagctctat
      721 gagcatgctg tagcccattc tggtctaccc gataacgtca agcacgcgtc cggtccgagc
      781 gcttccggac ctaggcccaa gaagaagcgc aaggtggcgc ccgggctcga gccatctgct
      841 ggagacatga gagctgccaa cctttggcca agcccgctca tgatcaaacg ctctaagaag
      901 aacagcctgg ccttgtccct gacggccgac cagatggtca gtgccttgtt ggatgctgag
      961 ccccccatac tctattccga gtatgatcct accagaccct tcagtgaagc ttcgatgatg
     1021 ggcttactga ccaacctggc agacagggag ctggttcaca tgatcaactg ggcgaagagg
     1081 gtgccaggct ttgtggattt gaccctccat gatcaggtcc accttctaga atgtgcctgg
     1141 ctagagatcc tgatgattgg tctcgtctgg cgctccatgg agcacccagt gaagctactg
     1201 tttgctccta acttgctctt ggacaggaac cagggaaaat gtgtagaggg catggtggag
     1261 atcttcgaca tgctgctggc tacatcatct cggttccgca tgatgaatct gcagggagag
     1321 gagtttgtgt gcctcaaatc tattattttg cttaattctg gagtgtacac atttctgtcc
     1381 agcaccctga agtctctgga agagaaggac catatccacc gagtcctgga caagatcaca
     1441 gacactttga tccacctgat ggccaaggca ggcctgaccc tgcagcagca gcaccagcgg
     1501 ctggcccagc tcctcctcat cctctcccac atcaggcaca tgagtaacaa aggcatggag
     1561 catctgtaca gcatgaagtg caagaacgtg gtgcccctct atgacctgct gctggaggcg
     1621 gcggacgccc accgcctaca tgcgcccact agccgtggag gggcatccgt ggaggagacg
     1681 gaccaaagcc acttggccac tgcgggctct acttcatcgc attccttgca aaagtattac
     1741 atcacggggg aggcagaggg tttccctgcc acagcttgat gaagatctga gggcccgttt
     1801 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1861 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1921 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1981 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     2041 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
     2161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4681 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4741 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
     4801 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     4861 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     4921 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
     4981 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     5041 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     5101 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     5161 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     5221 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     5281 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     5341 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     5401 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     5461 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     5521 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
     5581 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     5641 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     5701 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga ac
//
