LOCUS Exported 8166 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For mamalian expression of Ecdysone receptor-based inducible transgene expression switch.. ACCESSION . VERSION . KEYWORDS pEUI(+) 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8166 /organism="synthetic DNA construct" /mol_type="other DNA" 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" primer_bind 3478..3497 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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" promoter complement(5907..5924) /label=T7 promoter /note="promoter for bacteriophage T7 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 6437..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 //