LOCUS       sequence-241261-        6282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the marked mRNA under the control of substitutional 
            promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6282)
  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 6282)
  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: pEC005
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..23
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    140..161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(176..181,182..199,200..206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    214..230
                     /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)."
     primer_bind     219..241
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     238..254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     238..254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        275..293
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1261..1279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1282..1300
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1302..1320
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1368..1386
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2123..2141)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2333..2352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2334..2352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2359..2376)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2359..2375)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2368..2390)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2516..2971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2603..2622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2813..2834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3102..3905)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3906..4121)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4145..4164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4264..4286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4324..4342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4410..4514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4515..4583,4584..5375)
                     /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     complement(4733..4752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5546..6134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6035..6054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
       61 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      121 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      181 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      241 gaaacagcta tgaccatgat tacgccaagc tcgaaattaa ccctcactaa agggaacaaa
      301 agctggtacc atcatagaga aatcatatga aaaatcaaat acaactctgg gcttggagga
      361 agtcattata agatccggtt acatgggtgt ttgtattgat aacattccct cccaatatag
      421 ctcttacaat aatatgacta cattctccaa ttcaatttgc tatgcaagaa agaatttaag
      481 ctcggttccc ttatacagag acttggaaat tcaactctca aatattgcat cttccagctc
      541 caaaacccaa tcaagcgttg tcttgaacat tttgaagtta gcccaattaa cttcggttaa
      601 tgttatacat ccatatgtct tgatggcaac tgtaatccta acaatcctaa tgtttctatt
      661 tattctgtat gtaactgtcc ctaagttacc attcaaacta gcggttaata aatttctact
      721 gttgctaagt tcaactatag ttttgacatg gggtattggt gcgatgtgga ctcatgtagg
      781 gataaatgca agttacagat tggttccatc atcaagtatg aatataatca ctgtcaagaa
      841 gggcaagaag gcagcagtaa tggcttggtt tagctttgca ttcctccttt tagacagtgt
      901 ggttttatgg ttaatatttt taagggatag gaaaagcttg aaagatgaaa ttgataatgt
      961 tccatgtgcc caaaataggt acaataacta ctcttcggat tcatctacat tgcattccaa
     1021 agtttagctc gagggtcaga ttcattatct acaaagactg taaaaagaaa gtggaatatt
     1081 cattcatatc atattttttc tattaactgc ctggtttctt ttaaattttt tattggttgt
     1141 cgacttgaac ggagtgacaa tatatatata tatatattta ataatgacat cattatctgt
     1201 aaatctgatt cttaatgcta ttctagttat gtaagagtgg tcccctagga tattgaagta
     1261 tccctatcag tgatagagaa gtccctatca gtgatagaga atccctatca gtgatagaga
     1321 tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgccttcc ctatcagtga
     1381 tagagaaaca ataaagattc tacaatacta gcttttatgg ttatgaagag gaaaaattgg
     1441 cagtaacctg gccccacaaa ccttcaaatt aacgaatcaa attaacaacc ataggatgat
     1501 aatgcgatta gttttttagc cttatttctg gggtaattaa tcagcgaagc gatgattttt
     1561 gatctattaa cagatatata aatggaaaag ctgcagatct aatatagcgt agtaacatac
     1621 tgaaaagaga agcacagaaa aaataatcat agaagtgtta ggacatttga gggaacatat
     1681 ttcaacaata aacataatca ttgagtaaaa tggagctgat actaaacagc ctcatttctg
     1741 atgatttaac agaggaacaa aagcgcttga gccttgattt cctacaggat atcctacaat
     1801 ctaacacgaa agactatgaa agttattttt catctagggc tgttcctggt agcataactg
     1861 aagacatagc agaaattgat gctgaattat ctgcccttga tagaaagata aggaaaacac
     1921 tactagataa tacgtctcag attatcggaa atattcttga aaacgatgat agggctcaat
     1981 tggatgatat agccaagtct ttggagcaat tatgggaatt ggatactaat attaataagg
     2041 cagccgatcg aaacgttaca aatgacgata taaacaatga aagaattctg tacaaatagg
     2101 gccgcatcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     2161 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat tttcttatag
     2221 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacggac
     2281 gcgtgtacgc atctagctag agcggccgcc accgcggtgg agctccaatt cgccctatag
     2341 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2401 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2461 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2941 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3001 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     3061 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     3121 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     3181 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     3241 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     3301 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     3361 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     3421 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     3481 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     3541 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     3601 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     3661 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     3721 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     3781 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     3841 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     3901 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     3961 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     4021 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     4081 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc
     4141 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4201 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4261 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4321 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4381 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4441 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4501 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4561 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4621 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4681 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4741 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4801 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4861 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4921 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4981 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5041 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5101 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5341 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5401 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5461 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5521 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5581 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5641 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5701 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5761 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5821 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5881 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5941 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6001 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6061 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6181 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6241 cctttgagtg agctgatacc gctcgccgca gccgaacgac cg
//
