LOCUS       sequence-365706-        6349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclofen-inducible P38 activation via mammalian cell transfection or
            mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  TITLE     Vriz lab cyclofen-inducible ERT2 plasmids
REFERENCE   2  (bases 1 to 6349)
  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: pCSmp38ERT(#143)
FEATURES             Location/Qualifiers
     source          1..6349
                     /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             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             142..171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             181..210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             220..249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             259..288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1435..2370
                     /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             1483..2376
                     /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    2401..2535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2425..2444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2479..2498
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2648..2668)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2648..2666)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2687..2703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2687..2703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2700..2722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2711..2727
                     /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(2735..2741,2742..2759,2760..2765))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2780..2801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2918..2935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3089..3677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3848..4639,4640..4708))
                     /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     4471..4490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4709..4813)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4839..5294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4976..4997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5188..5207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5365..6349
                     /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 tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggcgcagaaa gaaagaccca ctttctatcg acaggaggtc
      361 aacaagacga tttgggaggt ccccgtgcag tatcagaact tgtctcccgt gggctccggt
      421 gcatacggat ctgtttgctc tgcgtttgat gcaaagacag gatttaaagt tgctgtgaag
      481 aagctctcga ggccatttca gtccatcatc catgccaagc ggacctacag ggagctgagg
      541 ctactcaagc acatgagaca tgagaatgtg atcggcttgc tggatgtttt tacacctgcc
      601 acaagtctga aggagtttaa tgatgtgtac ctggtgactc accttatggg cgctgacctg
      661 aataacatcg taaagtgtca gaaactgacc gacgatcacg tccagttcct catttaccag
      721 atactgagag gactaaagta catccattca gctgacataa ttcacaggga tttgaaaccc
      781 agtaatctgg cagtaaatga agactgtgaa ctcaagattc ttgactttgg cttggctcga
      841 cacacagatg atgagatgac tgggtatgtg gccacaaggt ggtatcgtgc ccctgagatc
      901 atgctcaact ggatgcacta caacgtgacg gtggacattt ggtcagtggg ctgcatcatg
      961 gctgagctgc tgacgggcag gactctgttc cctggcaccg atcatataaa ccagcttcag
     1021 cagataatgc gacttacagg aactcctcct tcctctctaa taagcaggat gcctagccat
     1081 gaggctcgta cttacatcag ctcacttcct cagatgccca agaggaattt tgcagatgtg
     1141 tttatcgggg caaatccaca agctgtggac ctcttggaga aaatgctggt tttggacact
     1201 gataagcgga ttacagcagc agaggctctg gcacatccat actttgccca atatcatgac
     1261 ccggatgatg aacccgaagc agagcctttt gaccagagct ttgagagccg agagctggat
     1321 attgaggagt ggaaacgtca gacatacgag gagatgatct cctttgagcc tccagtcttt
     1381 gatgtggatg aaatggagtc cggtccgtcc ggagcgcccg ggctcgagcc atctgctgga
     1441 gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc taagaagaac
     1501 agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga tgctgagccc
     1561 cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc gatgatgggc
     1621 ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc gaagagggtg
     1681 ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg tgcctggcta
     1741 gagatcctga tgattggtct cgtctggcgc tccatggagc acccagtgaa gctactgttt
     1801 gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat ggtggagatc
     1861 ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca gggagaggag
     1921 tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt tctgtccagc
     1981 accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa gatcacagac
     2041 actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca ccagcggctg
     2101 gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg catggagcat
     2161 ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct ggaggcggcg
     2221 gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga ggagacggac
     2281 caaagccact tggccactgc gggctctact tcatcgcatt ccttgcaaaa gtattacatc
     2341 acgggggagg cagagggttt ccctgccaca gcttgatgaa gatctgaggg cccgtttgta
     2401 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2461 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2521 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2581 gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct
     2641 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     2701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt
     2761 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2821 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2881 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4801 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     4861 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4921 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4981 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5041 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5101 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5161 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5221 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5281 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5341 gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca
     5401 tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg
     5461 tgccaattca atatggtgga tctggacccc agccaattca atatggcgga cttggcacca
     5521 tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg
     5581 gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg
     5641 gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg caatatccaa
     5701 tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc
     5761 atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt
     5821 cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca
     5881 atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat
     5941 ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga
     6001 cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg
     6061 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
     6121 tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca
     6181 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
     6241 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
     6301 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
