LOCUS       sequence-210121-        8166 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For mamalian expression of Ecdysone receptor-based inducible 
            transgene expression switch..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8166)
  AUTHORS   Lee S, Sohn KC, Choi DK, Won M, Park KA, Ju SK, Kang K, Bae YK, Hur 
            GM, Ro H
  TITLE     Ecdysone Receptor-based Singular Gene Switches for Regulated 
            Transgene Expression in Cells and Adult Rodent Tissues.
  JOURNAL   Mol Ther Nucleic Acids. 2016 Sep 27;5(9):e367. doi: 
            10.1038/mtna.2016.74.
  PUBMED    27673563
REFERENCE   2  (bases 1 to 8166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Nucleic Acids."; date: "2016-09-27"; volume: "5(9):e367. doi"; 
            pages: " 10.1038/mtna.2016.74"
COMMENT     Sequence Label: pEUI(+)
FEATURES             Location/Qualifiers
     source          1..8166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     44..61
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        177..556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        557..760
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     711..731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        805..823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             843..1280
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLQSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTV"
     promoter        complement(2385..2403)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2423..2440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2429..2653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2699..3127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2786..2805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2996..3017
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3193..3212)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3236..3565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3416..3551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3658..4257
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(3658..3677)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4154..4174
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4487..4621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    5376..5396
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        5398..5416
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5441..5457
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5442..5458
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5797..5815
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    5929..6063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6176..6196)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6176..6194)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6246..6264)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6332..6436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6437..6505,6506..7297)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6655..6674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7468..8056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7957..7976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
       61 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      121 ccgcatagtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      181 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      241 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      301 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      361 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      421 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      481 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      541 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      601 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      661 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      721 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      781 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttatcg
      841 atatgaagct acagtcttct atcgaacaag catgcgatat ttgccgactt aaaaagctca
      901 agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct
      961 actctcccaa aaccaaaagg tctccgctga ctagggcaca tctgacagaa gtggaatcaa
     1021 ggctagaaag actggaacag ctatttctac tgatttttcc tcgagaagac cttgacatga
     1081 ttttgaaaat ggattcttta caggatataa aagcattgtt aacaggatta tttgtacaag
     1141 ataatgtgaa taaagatgcc gtcacagata gattggcttc agtggagact gatatgcctc
     1201 taacattgag acagcataga ataagtgcga catcatcatc ggaagagagt agtaacaaag
     1261 gtcaaagaca gttgactgta gagttcctgg atatggccga cttcgagttt gagcagatgt
     1321 ttaccgatgc ccttggaatt gacgagtacg gtgggctgca gatgaggcct gaatgtgtca
     1381 tacaggagcc cagtaaaaat aaagacaggc aaagacaaaa gaaagacaaa ggaatattat
     1441 tacctgttag tacgaccaca gtcgaagacc acatgccccc gatcatgcaa tgtgatccac
     1501 ctccgcccga ggccgccagg attcacgaag tcgtcccgag gtatctttcg gagaagctga
     1561 tggagcagaa caggcagaag aacataccac cattgtcggc gaatcagaag tctctgatcg
     1621 cgaggctcgt gtggtaccag gagggatatg agcagccctc cgacgaggat ctcaaaagag
     1681 taacgcagac ttggcagtcg gatgaagagg acgaggaatc cgatctaccc ttccgccaga
     1741 tcacggagat gacgatctta acggtccagt tgatcgtcga gttcgccaag ggtctaccgg
     1801 gcttttcgaa gatatcacag tctgatcaaa tcaccttatt aaaagcctcg tccagcgagg
     1861 tgatgatgct gcgggtggcg aggcgatacg acgccgcgtc cgacagcatt ctgttcgcca
     1921 acaacaaggc gtacacgcgc gacaactacc gcaaggcggg catggccgag gtcatcgaag
     1981 acctcctaca cttctgccgg tgcatgttcg cgatgggcat ggacaatgtg cactttgcac
     2041 tgctcacggc catcgttata ttctcagatc ggcccgggct cgagcagccg tcgctggtag
     2101 aagagatcca gagatactac ctgaacacgt tgcgaattta catcatcaac cagaacagcg
     2161 cgtcgtcgcg ctgcgccgtg atctacggca ggatcctgag cgtgctgacc gagctacgca
     2221 cgctcggcac gcaaaactcc aacatgtgca tctcgctgaa gctgaagaac aggaagctgc
     2281 cgccgttcct cgaggagatc tgggacgtgg cggaggtggc cacgacgcat cccacggtgc
     2341 tgccgcccac caacccggtg gtgctatagt ctagagggcc ctattctata gtgtcaccta
     2401 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2461 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2521 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2581 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc cggggatgcg
     2641 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2701 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2761 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2821 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2881 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2941 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3001 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3061 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3121 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctcctatg
     3181 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagctgt ggaatgtgtg
     3241 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3301 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3361 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3421 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3481 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3541 ttttggaggc ctaggctttt gcaaaaagct tgcatgcctg caggtcggcc gccacgaccg
     3601 gtgccgccac catcccctga cccacgcccc tgacccctca caaggagacg accttccatg
     3661 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc
     3721 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc
     3781 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     3841 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     3901 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     3961 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     4021 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     4081 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     4141 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     4201 gtcgagtgcc cgaaggaccg cgcgacctgg tgcatgaccc gcaagcccgg tgcctgacgc
     4261 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgg ctccgaccga
     4321 agccgacccg ggcggccccg ccgaccccgc acccgccccc gaggcccacc gactctagag
     4381 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4441 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4501 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4561 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4621 ccccaggaag ctcctctgtg tcctcataaa ccctaacctc ctctacttga gaggacattc
     4681 caatcatagg ctgcccatcc accctctgtg tcctcctgtt aattaggtca cttaacaaaa
     4741 aggaaattgg gtaggggttt ttcacagacc gctttctaag ggtaatttta aaatatctgg
     4801 gaagtccctt ccactgctgt gttccagaag tgttggtaaa cagcccacaa atgtcaacag
     4861 cagaaacata caagctgtca gctttgcaca agggcccaac accctgctca tcaagaagca
     4921 ctgtggttgc tgtgttagta atgtgcaaaa caggaggcac attttcccca cctgtgtagg
     4981 ttccaaaata tctagtgttt tcatttttac ttggatcagg aacccagcac tccactggat
     5041 aagcattatc cttatccaaa acagccttgt ggtcagtgtt catctgctga ctgtcaactg
     5101 tagcattttt tggggttaca gtttgagcag gatatttggt cctgtagttt gctaacacac
     5161 cctgcagctc caaaggttcc ccaccaacag caaaaaaatg aaaatttgac ccttgaatgg
     5221 gttttccagc accattttca tgagtttttt gtgtccctga atgcaagttt aacatagcag
     5281 ttaccccaat aacctcagtt ttaacagtaa cagcttccca catcaaaata tttccacagg
     5341 ttaagtcctc atttaaatta ggcaaaggaa ttgctcaact ttgtatagaa aagttggtaa
     5401 tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcga cggtatcgat
     5461 aagcttaggc ctccaaggcg gagtactgtc ctccgggctg gcggagtact gtcctccggc
     5521 aaggtcggag tactgtcctc cgacactaga ggtcggagta ctgtcctccg acgcaaggcg
     5581 gagtactgtc ctccgggctg cggagtactg tcctccggca aggtcggagt actgtcctcc
     5641 gacactagag gtcggagtac tgtcctccga cgcaaggcgg agtactgtcc tccgggctgg
     5701 cggagtactg tcctccggca agggtcgact ctagagggta tataatggat cccatcgcgt
     5761 ctcagcctca ctttgagctc ctccacacgg agcttgattt aggtgacact atagaataca
     5821 agctacttgt tctttttgca ggatcccatc gattcgaatt cacgtgctag cgcttaagaa
     5881 attcaaggcc tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
     5941 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     6001 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     6061 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     6121 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     6181 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgttaatt cttgaagacg
     6241 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6301 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6361 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6421 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6481 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6541 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6601 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6661 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6721 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6781 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6841 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6901 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6961 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     7021 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7081 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7141 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7201 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7261 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8161 cgcctt
//
