LOCUS       sequence-365942-        6523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible mouse Pax6 activation via mammalian cell 
            transfection or mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6523)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 6523)
  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: pCSThaP6ERT(#184)
FEATURES             Location/Qualifiers
     source          1..6523
                     /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     140..159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        140..158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           191..233
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             274..300
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1576..1596
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1609..2544
                     /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             1657..2550
                     /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    2575..2709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2599..2618)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2653..2672
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2822..2842)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2822..2840)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2861..2877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2861..2877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2874..2896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2885..2901
                     /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(2909..2915,2916..2933,2934..2939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2954..2975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3092..3109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3263..3851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3343..3362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4022..4813,4814..4882))
                     /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     4645..4664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4883..4987)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5013..5468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5150..5171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5362..5381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5539..6523
                     /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 ttcctctctg gctaactaga gaacccactg
      121 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagccta
      181 agctagcttg cttgttcttt ttgcagaagc tcagaataaa cgctcaactt tggcagatac
      241 cagatccgga ggctgccaac cagctccatg cattacccat acgatgttcc agattacgcc
      301 ggcatgcaga acagtcacag cggagtgaat cagcttggtg gtgtctttgt caacgggcgg
      361 ccactgccgg actccacccg gcagaagatc gtagagctag ctcacagcgg ggcccggccg
      421 tgcgacattt cccgaattct gcaggtatcc aacggttgtg tgagtaaaat tctgggcagg
      481 tattacgaga ctggctccat cagacccagg gcaatcggag ggagtaagcc aagagtggcg
      541 actccagaag ttgtaagcaa aatagcccag tataaacggg agtgcccttc catctttgct
      601 tgggaaatcc gagacagatt attatccgag ggggtctgta ccaacgataa catacccagt
      661 gtgtcatcaa taaacagagt tcttcgcaac ctggctagcg aaaagcaaca gatgggcgca
      721 gacggcatgt atgataaact aaggatgttg aacgggcaga ccggaagctg gggcacacgc
      781 cctggttggt atcccgggac ttcagtacca gggcaaccca cgcaagatgg ctgccagcaa
      841 caggaaggag ggggagagaa caccaactcc atcagttcta acggagaaga ctcggatgaa
      901 gctcagatgc gacttcagct gaagcggaag ctgcaaagaa atagaacatc ttttacccaa
      961 gagcagattg aggctctgga gaaagagttt gagaggaccc attatccaga tgtgtttgcc
     1021 cgggaaagac tagcagccaa aatagatcta cctgaagcaa gaatacaggt atggttttct
     1081 aatcgaaggg ccaaatggag aagagaagag aaactgagga accagagaag acaggccagc
     1141 aacactccta gtcacattcc tatcagcagc agcttcagta ccagtgtcta ccagccaatc
     1201 ccacagccca ccacacctgt ctcctccttc acatcaggtt ccatgttggg ccgaacagac
     1261 accgccctca ccaacacgta cagtgctttg ccacccatgc ccagcttcac catggcaaac
     1321 aacctgccta tgcaaccccc agtccccagt cagacctcct cgtactcgtg catgctgccc
     1381 accagcccgt cagtgaatgg gcggagttat gatacctaca cccctccgca catgcaaaca
     1441 cacatgaaca gtcagcccat gggcacctcg gggaccactt caacaggact catttcacct
     1501 ggagtgtcag ttcccgtcca agttcccggg agtgaacctg acatgtctca gtactggcct
     1561 cgattacagg cgcccgagaa cctgtacttc caggggctcg agccatctgc tggagacatg
     1621 agagctgcca acctttggcc aagcccgctc atgatcaaac gctctaagaa gaacagcctg
     1681 gccttgtccc tgacggccga ccagatggtc agtgccttgt tggatgctga gccccccata
     1741 ctctattccg agtatgatcc taccagaccc ttcagtgaag cttcgatgat gggcttactg
     1801 accaacctgg cagacaggga gctggttcac atgatcaact gggcgaagag ggtgccaggc
     1861 tttgtggatt tgaccctcca tgatcaggtc caccttctag aatgtgcctg gctagagatc
     1921 ctgatgattg gtctcgtctg gcgctccatg gagcacccag tgaagctact gtttgctcct
     1981 aacttgctct tggacaggaa ccagggaaaa tgtgtagagg gcatggtgga gatcttcgac
     2041 atgctgctgg ctacatcatc tcggttccgc atgatgaatc tgcagggaga ggagtttgtg
     2101 tgcctcaaat ctattatttt gcttaattct ggagtgtaca catttctgtc cagcaccctg
     2161 aagtctctgg aagagaagga ccatatccac cgagtcctgg acaagatcac agacactttg
     2221 atccacctga tggccaaggc aggcctgacc ctgcagcagc agcaccagcg gctggcccag
     2281 ctcctcctca tcctctccca catcaggcac atgagtaaca aaggcatgga gcatctgtac
     2341 agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc tgctggaggc ggcggacgcc
     2401 caccgcctac atgcgcccac tagccgtgga ggggcatccg tggaggagac ggaccaaagc
     2461 cacttggcca ctgcgggctc tacttcatcg cattccttgc aaaagtatta catcacgggg
     2521 gaggcagagg gtttccctgc cacagcttga tgaagatctg agggcccgtt tgtagatcca
     2581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2701 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2761 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     2821 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2881 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     2941 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3001 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3061 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3961 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4021 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4141 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4621 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4981 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5041 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5101 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5161 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5221 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5281 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5341 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5401 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5461 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5521 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     5581 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     5641 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     5701 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     5761 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     5821 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     5881 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     5941 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     6001 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     6061 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     6121 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     6181 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     6241 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     6301 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     6361 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     6421 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     6481 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
