LOCUS       sequence-365692-        5595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible GAL4 activation via mammalian cell transfection 
            or mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5595)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 5595)
  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: pCSTG4FER3(#190)
FEATURES             Location/Qualifiers
     source          1..5595
                     /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"
     primer_bind     84..103
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        84..102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           121..163
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             172..615
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MGKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKT
                     KRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVN
                     KDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             703..723
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             736..1671
                     /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             784..1677
                     /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    1702..1836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1726..1745)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1780..1799
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1933..1949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1933..1949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1946..1968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1957..1973
                     /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(1981..1987,1988..2005,2006..2011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2026..2047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2164..2181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2415..2434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3094..3885,3886..3954))
                     /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     3717..3736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3955..4059)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4085..4540)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4222..4243)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4434..4453
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4611..5595
                     /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 atctaatacg actcactata gggcgcgcta gcctaggttg
      121 cttgttcttt ttgcagaagc tcagaataaa cgctcaactt tggcagatac catggggaag
      181 ctactgtctt ctatcgaaca agcctgcgat atttgccgac ttaaaaagct gaagtgctcc
      241 aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc
      301 aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa
      361 agactggaac agctatttct actgattttt ccccgcgaag accttgacat gattttgaaa
      421 atggattctt tacaggatat aaaagcactg ctaacaggat tattcgtaca agataatgtg
      481 aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg
      541 agacagcata gaataagtgc gacatcatca tcggaggaaa gtagtaacaa aggtcaaaga
      601 cagctcactg tatcgcccgc cgacgcactg gacgacttcg acctggacat gctgcctgct
      661 gatgctctcg atgactttga tctcgatatg ctgcccgggc cagagaacct gtacttccag
      721 gggctcgagc cctctgccgg agacatgaga gctgcaaacc tttggccgag cccgctgatg
      781 attaaacgct ctaagaagaa cagcctggcc ttgtccctga ccgccgacca gatggtcagt
      841 gccttgttgg atgccgagcc ccccatactc tattccgagt atgatcctac cagacccttc
      901 agtgaggctt cgatgatggg cttactgacc aacctggcag acagggagct ggttcacatg
      961 attaactggg ccaagagggt gccaggcttt gtggatttga ccctccacga tcaggtccac
     1021 ctcctagaat gtgcctggct agagatcctg atgattggtc tcgtctggcg cagcatggag
     1081 cacccagtga agctactgtt tgctcctaac ttgctcttgg acaggaacca gggaaaatgt
     1141 gtagagggca tggtggagat cttcgacatg ctgctggcta catcatctcg gttccgcatg
     1201 atgaatctgc agggagagga gtttgtgtgc ctcaaatcta ttattttgct taattctgga
     1261 gtgtacacat ttctgtccag caccctgaag tctctggaag agaaggacca tatccaccga
     1321 gtcctggaca agatcacaga cactttgatc cacctgatgg ccaaggcagg cctgaccctg
     1381 cagcagcagc accagcggct ggcccagctc ctcctcatcc tctcccacat caggcacatg
     1441 agtaacaaag gcatggagca tctgtacagc atgaagtgca agaacgtggt gcccctctat
     1501 gacctgctgc tggaggcggc ggacgcccac cgcctacatg cgcccactag ccgtggaggg
     1561 gcatccgtgg aggagacgga ccaaagccac ttggccactg cgggctctac ttcatcgcat
     1621 tccttgcaaa agtattacat cacgggggag gcagagggtt tccctgccac agcttgatga
     1681 agatctgagg gcccgtttgt agatccagac atgataagat acattgatga gtttggacaa
     1741 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1801 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1861 atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggtcgcgctt
     1921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1981 caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2641 tcttgagtcc aacccagtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3061 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3181 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3301 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3421 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3541 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3601 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3721 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3781 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3961 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4021 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4081 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4141 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4201 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4261 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4321 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4381 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4441 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4501 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     4561 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc
     4621 aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc
     4681 aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc
     4741 aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc
     4801 aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc
     4861 caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc
     4921 aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt
     4981 tcaatactat gtattggccc tatgccatat agtattccat atatgggttt tcctattgac
     5041 gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc
     5101 ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc
     5161 atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat
     5221 ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg
     5281 gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa
     5341 atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat
     5401 tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc
     5461 cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg
     5521 ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat
     5581 gctcgtttag ggaac
//
