LOCUS       sequence-365952-        5086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to express cyclofen-inducible fusions in mammalian cells or fish 
            eggs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5086)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 5086)
  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: pCSnERT(#186)
FEATURES             Location/Qualifiers
     source          1..5086
                     /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             130..150
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             172..1107
                     /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             220..1113
                     /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    1138..1272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1162..1181)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1216..1235
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1385..1405)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1385..1403)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1424..1440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1424..1440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1437..1459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1448..1464
                     /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(1472..1478,1479..1496,1497..1502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1517..1538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1655..1672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1826..2414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1906..1925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2585..3376,3377..3445))
                     /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     3208..3227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3446..3550)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3576..4031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3713..3734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3925..3944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4102..5086
                     /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 atcctaggct agcaccggcg gcatggacga gctgctgtac
      121 aaatcgatcc ccaagaagaa gcgcaaggtg gcgcccgggc tcgagccatc tgctggagac
      181 atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc
      241 ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc
      301 atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta
      361 ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca
      421 ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag
      481 atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagtgaagct actgtttgct
      541 cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc
      601 gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt
      661 gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc
      721 ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact
      781 ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc
      841 cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg
      901 tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga ggcggcggac
      961 gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa
     1021 agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg
     1081 ggggaggcag agggtttccc tgccacagct tgatgaagat ctgagggccc gtttgtagat
     1141 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1201 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1261 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1321 tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt
     1381 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     1441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta
     1501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3541 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     3601 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3661 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3721 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3781 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3841 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3901 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3961 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4021 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4081 ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc
     4141 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
     4201 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
     4261 caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg
     4321 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
     4381 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
     4441 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
     4501 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
     4561 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
     4621 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
     4681 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
     4741 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
     4801 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
     4861 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
     4921 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     4981 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     5041 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
