LOCUS Exported 7244 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses LexA-ER-B42 in yeast cells. ACCESSION . VERSION . KEYWORDS FRP718_PACT1(-1-520)-LexA-ER-haB42-TCYC1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7244) AUTHORS Ottoz DS, Rudolf F, Stelling J TITLE Inducible, tightly regulated and growth condition-independent transcription factor in Saccharomyces cerevisiae. JOURNAL Nucleic Acids Res. 2014 Jul 17. pii: gku616. PUBMED 25034689 REFERENCE 2 (bases 1 to 7244) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7244 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS 509..1168 /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" rep_origin complement(1429..1884) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1566..1587) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1778..1797 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2014..2036 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2028..2045 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2029..2045 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2052..2071 /label=T7 /note="T7 promoter, forward primer" promoter 2052..2070 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2709..3314 /codon_start=1 /gene="lexA" /product="E. coli DNA-binding protein that represses genes involved in the SOS response to DNA damage" /label=LexA /note="The N-terminal DNA-binding domain recognizes the 16-bp palindromic sequence 5'-CTGTATATATATACAG-3'." /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL" primer_bind 3252..3273 /label=LexA /note="LexA DNA binding domain, forward primer" CDS 3381..4271 /codon_start=1 /gene="ESR1" /product="estrogen nuclear receptor alpha ligand-binding domain" /label=ER-LBD /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL QKYYITGEAEGFPATV" CDS 4320..4556 /codon_start=1 /product="yeast transcriptional activator cretaed from E. coli genomic DNA fragments (Ma and Ptashne, 1987)" /label=B42 transcriptional activator /translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD" primer_bind complement(4589..4605) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4590..4606) /label=pBluescriptKS /note="For pBluescript vector" terminator 4633..4882 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(4649..4667) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" promoter complement(4994..5012) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(5033..5049) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5033..5049) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5046..5068) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5057..5073 /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(5081..5111) /label=lac promoter /note="promoter for the E. coli lac operon" primer_bind complement(5264..5281) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5435..6023) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6194..7054) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6817..6836 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7055..7159) /gene="bla" /label=AmpR promoter primer_bind 7217..7235 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta tcgtttgaac 241 acggcattag tcagggaagt cataacacag tcctttcccg caattttctt tttctattac 301 tcttggcctc ctctagtaca ctctatattt ttttatgcct cttaattaag gtaatgattt 361 tcattttttt ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc 421 acataatgaa ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa 481 tgagcaggca agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat 541 tacaaatgaa accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga 601 gcactcgatc ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca 661 agtgattaac gtccacacag gtatagggtt tctggaccac atgatacatg ctctggccaa 721 gcattccggc tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca 781 caccactgaa gactgcggga ttgctctcgg tcaggctttt aaagaggccc tactggcgcg 841 tggagtaaaa aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt 901 ggtagatctt tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt 961 aggagatctc tcttgcgaga tgatcccgca ttttcttgaa agttttgcag aggctagcag 1021 aattaccctc cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc 1081 gttcaaggct cttgcggttg ccataagaga agccacctcg cccaatggaa ccaacgatgt 1141 tccctccacc aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga 1201 tatatataca tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt 1261 atgatactga agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc 1321 gctttccttt tttctttttg ctttttcttt ttttttctct tgaactcgac ggatcatatg 1381 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg 1441 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 1501 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 1561 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 1621 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 1681 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 1741 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 1801 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 1861 ttaatgcgcc gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg 1921 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg 1981 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 2041 ccagtgaatt gtaatacgac tcactatagg gcgaattgga gaaacagcgg ccgcctatag 2101 ggcgaattgg agctccaccg cggtggcggc cgctctagag aagcgggtaa gctgccacag 2161 caattaatgc acaacattta acctacattc ttccttatcg gatcctcaaa acccttaaaa 2221 acatatgcct caccctaaca tattttccaa ttaaccctca atatttctct gtcacccggc 2281 ctctattttc cattttcttc tttacccgcc acgcgttttt ttctttcaaa tttttttctt 2341 ccttcttctt tttcttccac gtcctcttgc ataaataaat aaaccgtttt gaaaccaaac 2401 tcgcctctct ctctcctttt tgaaatattt ttgggtttgt ttgatccttt ccttcccaat 2461 ctctcttgtt taatatatat tcatttatat cacgctctct ttttatcttc ctttttttcc 2521 tctctcttgt attcttcctt cccctttcta ctcaaaccaa gaagaaaaag aaaaggtcaa 2581 tctttgttaa agaataggat cttctactac atcagctttt agatttttca cgcttactgc 2641 ttttttcttc ccaagatcga aaatttactg aattaacaac tagtggatcc cccgggctgc 2701 aggaattcat gaaagcgtta acggccaggc aacaagaggt gtttgatctc atccgtgatc 2761 acatcagcca gacaggtatg ccgccgacgc gtgcggaaat cgcgcagcgt ttggggttcc 2821 gttccccaaa cgcggctgaa gaacatctga aggcgctggc acgcaaaggc gttattgaaa 2881 ttgtttccgg cgcatcacgc gggattcgtc tgttgcagga agaggaagaa gggttgccgc 2941 tggtaggtcg tgtggctgcc ggtgaaccac ttctggcgca acagcatatt gaaggtcatt 3001 atcaggtcga tccttcctta ttcaagccga atgctgattt cctgctgcgc gtcagcggga 3061 tgtcgatgaa agacatcggc attatggatg gtgacttgct ggcagtgcat aaaactcagg 3121 atgtacgtaa cggtcaggtc gttgtcgcac gtattgatga cgaggttacc gttaagcgcc 3181 tgaaaaaaca gggcaataaa gtcgaactgt tgccagaaaa tagcgagttt aaaccaattg 3241 tcgtagatct tcgtcagcag agcttcacca ttgaagggct ggcggttggg gttattcgca 3301 acggcgactg gctgggttct aaggatatct ctgctggaga catgagagct gccaaccttt 3361 ggccaagccc gctcatgatc aaacgctcta agaagaacag cctggccttg tccctgacgg 3421 ccgaccagat ggtcagtgcc ttgttggatg ctgagccccc catactctat tccgagtatg 3481 atcctaccag acccttcagt gaagcttcga tgatgggctt actgaccaac ctggcagaca 3541 gggagctggt tcacatgatc aactgggcga agagggtgcc aggctttgtg gatttgaccc 3601 tccatgatca ggtccacctt ctagaatgtg cctggttaga gatcctgatg attggtctcg 3661 tctggcgctc catggagcac ccagtgaagc tactgtttgc tcctaacttg ctcttggaca 3721 ggaaccaggg aaaatgtgta gagggcatgg tggagatctt cgacatgctg ctggctacat 3781 catctcggtt ccgcatgatg aatctgcagg gagaggagtt tgtgtgcctc aaatctatta 3841 ttttgcttaa ttctggagtg tacacatttc tgtccagcac cctgaagtct ctggaagaga 3901 aggaccatat ccaccgagtc ctggacaaga tcacagacac tttgatccac ctgatggcca 3961 aggcaggcct gaccctgcag cagcagcacc agcggctggc ccagctcctc ctcatcctct 4021 cccacatcag gcacatgagt aacaaaggca tggagcatct gtacagcatg aagtgcaaga 4081 acgtggtgcc cctctatgac ctgctgctgg agatgctgga cgcccaccgc ctacatgcgc 4141 ccactagccg tggaggggca tccgtggagg agacggacca aagccacttg gccactgcgg 4201 gctctacttc atcgcattcc ttgcaaaagt attacatcac gggggaggca gagggtttcc 4261 ctgccacagt cgatatcatg caggatctgc cgggcaacga taacagcacc gcgggcggta 4321 tcaataaaga catcgaggag tgcaatgcca tcattgagca gtttatcgac tacctgcgca 4381 ccggacagga gatgccgatg gaaatggcgg atcaggcgat taacgtggtg ccgggcatga 4441 cgccgaaaac cattcttcac gccgggccgc cgatccagcc tgactggctg aaatcgaatg 4501 gttttcatga aattgaagcg gatgttaacg ataccagcct cttgctgagt ggagatgcct 4561 ccaagcttta aacgcgttta aagcttatcg ataccgtcga cctcgagaca ggcccctttt 4621 cctttgtcta tatcatgtaa ttagttatgt cacgcttaca ttcacgccct cctcccacat 4681 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt 4741 tttaatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct 4801 gtacaaacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg 4861 acgctcgaag gctttaattt gcaaggttcg cagtttacac tctcatcgtc gctctcatca 4921 tcgcttccgt tgttgttttc cttagtagcg tctgcttggt acccagcttc ggccgtgtac 4981 ccagcttttg ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc 5041 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatagga gccggaagca 5101 taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt gcgttgcgct 5161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 5221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 5281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 5341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 5401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctcggc ccccctgacg 5461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5521 accaggcgtt cccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct 5641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 5881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 6001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 6061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 6121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 6181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 6241 ttcgttcatc catagttgcc tgactgcccg tcgtgtagat aactacgata cgggagggct 6301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 6361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 6421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 6481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 6541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 6601 tgtgaaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 6661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 6721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 6781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 6841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 6961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 7021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 7081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 7141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc ttattatcat 7201 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc //