LOCUS       sequence-240650-        7839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the regulator tTA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7839)
  AUTHORS   Baudrimont A, Voegeli S, Viloria EC, Stritt F, Lenon M, Wada T, 
            Jaquet V, Becskei A
  TITLE     Multiplexed gene control reveals rapid mRNA turnover.
  JOURNAL   Sci Adv. 2017 Jul 12;3(7):e1700006. doi: 10.1126/sciadv.1700006. 
            eCollection 2017 Jul.
  PUBMED    28706991
REFERENCE   2  (bases 1 to 7839)
  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"; journalName: "Sci Adv."; 
            date: "2017-07-12"; volume: "3(7):e1700006. doi"; pages: " 
            10.1126/sciadv.1700006. eCollection 2017 Jul"
COMMENT     Sequence Label: pMG13
FEATURES             Location/Qualifiers
     source          1..7839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(616..1236,1237..1623)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MPRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(689..708)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1664..1682)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(1858..1876)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1897..1913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1897..1913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1910..1932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1921..1937
                     /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(1945..1951,1952..1969,1970..1975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1990..2011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2128..2145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2299..2887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2379..2398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3058..3849,3850..3918))
                     /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     3681..3700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3919..4023)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4091..4109
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4147..4169)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4269..4288
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(4782..5876)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5877..6284)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(6585..7040)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6722..6743)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6934..6953
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7166..7188
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7180..7197
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7181..7197
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7204..7223
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7204..7222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagaaaaagt gaaaaattat caggcaagaa aaagaaatta ccaagcctgc tctcactgta
       61 atgatcaagt tacataaatt tactatcgga ttagtgtgtc cctctgccac atttccatat
      121 ttggccttgg gtttttgccc tcatcttttt tttttcttcc tcctacctat ttataatttg
      181 tatatctgta ctttcctcga gcttttaatc ttcttcttaa acattttctt gtgtagtata
      241 ctttcacaca atttctttct tgattttttc ttctactact gagtctgcca gtcaaatgga
      301 tttctgagga aagaggacta tacccattag gaaacgaatt gcccgagtag tctcctctgc
      361 cgacttaaac caaccttttt ctatttctct tttcttttct ccctcttttt tctctgtact
      421 agcatccaaa agcaagcatc catccgagtc ccagtcgcaa tctcacatct ccaatttaac
      481 gtatccattg catttcctca ttcggtttaa ctcctctgca tttcttttct gacccatagc
      541 atttcttaca ttccattgca tctccctttt actctcgttc aagacactga tttgatacgc
      601 tttctgtacg gatccatgcc tagattagat aaaagtaaag tgattaacag cgcattagag
      661 ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt
      721 gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta
      781 gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg
      841 caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat
      901 ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat
      961 caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc
     1021 gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa
     1081 gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa
     1141 ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc
     1201 ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa
     1261 aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga cgcccccgaa
     1321 gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac gcgcagactg
     1381 tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac
     1441 gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg
     1501 gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg
     1561 gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtggg
     1621 taggaattct gtacaaatag ggccgcatca tgtaattagt tatgtcacgc ttacattcac
     1681 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     1741 tccctattta ttttcttata gttatgttag tattaagaac gttatttata tttcaaattt
     1801 ttcttttttt tctgtacgga cgcgtgtacg catcgggccc ggtaccagct tttgttccct
     1861 ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2701 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2821 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4021 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4081 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4141 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4201 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4261 taactatgcg gcatcagagc agattgtact gagagtgcac catatcgact acgtcgttaa
     4321 ggccgtttct gacagagtaa aattcttgag ggaactttca ccattatggg aaatggttca
     4381 agaaggtatt gacttaaact ccatcaaatg gtcaggtcat tgagtgtttt ttatttgttg
     4441 tatttttttt tttttagaga aaatcctcca atatataaat taggaatcat agtttcatga
     4501 ttttctgtta cacctaactt tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa
     4561 gtgcaattct ttttccttat cacgttgagc cattagtatc aatttgctta cctgtattcc
     4621 tttacatcct cctttttctc cttcttgata aatgtatgta gattgcgtat atagtttcgt
     4681 ctaccctatg aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt
     4741 ataaagttta tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta
     4801 acttcttcgg cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca
     4861 gttctgatac ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca
     4921 ggcaagttca atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg
     4981 accttattct ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg
     5041 gtgttcttgt ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg
     5101 ataacggagg cttcatcgga gatgatatca ccaaacatgt tgctggtgat tataatacca
     5161 tttaggtggg ttgggttctt aactaggatc atggcggcag aatcaatcaa ttgatgttga
     5221 accttcaatg tagggaattc gttcttgatg gtttcctcca cagtttttct ccataatctt
     5281 gaagaggcca aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt
     5341 agggccatga aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca
     5401 ctatcccaag cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc
     5461 actaattctc tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat
     5521 aagtctaaaa gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt
     5581 tctttacgga tttttagtaa accttgttca ggtctaacac taccggtacc ccatttagga
     5641 ccacccacag cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct
     5701 ggaagtggaa cacctgtagc atcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg
     5761 aacttgacat tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg
     5821 atttcttgac caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga
     5881 atggtatatc cttgaaatat atatatatat attgctgaaa tgtaaaaggt aagaaaagtt
     5941 agaaagtaag acgattgcta accacctatt ggaaaaaaca ataggtcctt aaataatatt
     6001 gtcaacttca agtattgtga tgcaagcatt tagtcatgaa cgcttctcta ttctatatga
     6061 aaagccggtt ccggcgctct cacctttcct ttttctccca atttttcagt tgaaaaaggt
     6121 atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg
     6181 tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag
     6241 attcgaactc ttgcatctta cgatacctga gtattcccac agttaactgc ggtcaagata
     6301 tttcttgaat caggcgcctt agaccgctcg gccaaacaac caattacttg ttgagaaata
     6361 gagtataatt atcctataaa tataacgttt ttgaacacac atgaacaagg aagtacagga
     6421 caattgattt tgaagagaat gtggattttg atgtaattgt tgggattcca tttttaataa
     6481 ggcaataata ttaggtatgt agatatacta gaagttctcc tcgaccggtc gatatgcggt
     6541 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     6601 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     6661 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     6721 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     6781 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     6841 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6901 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6961 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     7021 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7081 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     7141 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     7201 ttgtaatacg actcactata gggcgaattg gagctccacc gcggtggcgg ccgaatgata
     7261 tacagccagc gtaagtacgt gtaaatagcc aagtaagaaa taatgatgtc ctagtgcagc
     7321 cacaatcaat ttcacttttt catatatatt tttccacctt attatcacaa ggtgcacctt
     7381 tatcgtgtgc cacggcggta aaaaactacc ttcaggggta cgatacattc tgtgctggcg
     7441 accacggggc tgacagagac acccgtagag gctacattac tgatttggga aatttcccaa
     7501 attggaaata tcacctcgtc gacgtgctgc ggtgcatggc tgttttaccc gtttaggaaa
     7561 aaactcggcg ggttttcttg acgggcaaat gtcgccattc gtctcgtttg aacgcttggc
     7621 ctgtaaaaca gctaactcat tcactatctc tatcctgcga ccgtgatgat accgaaagta
     7681 tgctgataca gatcggtccc tcatatcgcg atagggcttt ctgagctctc ctcccccttc
     7741 ttccctcccc agccaaagag gccggttttc cttctgggga agtgtcgcaa ccaaacgctg
     7801 tgctcaaacc ctaaattgtg aactttcaag aaaaaaaaa
//
