LOCUS       sequence-241015-        6269 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 6269)
  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 6269)
  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: pSV_207
FEATURES             Location/Qualifiers
     source          1..6269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    123..144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(159..164,165..182,183..189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..213
                     /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     202..224
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     221..237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     221..237
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        258..276
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1244..1262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1265..1283
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1285..1303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1351..1369
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2093..2111)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2303..2322)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2304..2322)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2329..2346)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2329..2345)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2338..2360)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2486..2941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2783..2804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3072..3875)
                     /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(3876..4091)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4115..4134)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4234..4256
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4294..4312)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4380..4484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4485..4553,4554..5345)
                     /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(4703..4722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5516..6104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6005..6024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6258..6269,1..6)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      181 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      241 gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt accatcatag
      301 agaaatcata tgaaaaatca aatacaactc tgggcttgga ggaagtcatt ataagatccg
      361 gttacatggg tgtttgtatt gataacattc cctcccaata tagctcttac aataatatga
      421 ctacattctc caattcaatt tgctatgcaa gaaagaattt aagctcggtt cccttataca
      481 gagacttgga aattcaactc tcaaatattg catcttccag ctccaaaacc caatcaagcg
      541 ttgtcttgaa cattttgaag ttagcccaat taacttcggt taatgttata catccatatg
      601 tcttgatggc aactgtaatc ctaacaatcc taatgtttct atttattctg tatgtaactg
      661 tccctaagtt accattcaaa ctagcggtta ataaatttct actgttgcta agttcaacta
      721 tagttttgac atggggtatt ggtgcgatgt ggactcatgt agggataaat gcaagttaca
      781 gattggttcc atcatcaagt atgaatataa tcactgtcaa gaagggcaag aaggcagcag
      841 taatggcttg gtttagcttt gcattcctcc ttttagacag tgtggtttta tggttaatat
      901 ttttaaggga taggaaaagc ttgaaagatg aaattgataa tgttccatgt gcccaaaata
      961 ggtacaataa ctactcttcg gattcatcta cattgcattc caaagtttag ctcgagggtc
     1021 agattcatta tctacaaaga ctgtaaaaag aaagtggaat attcattcat atcatatttt
     1081 ttctattaac tgcctggttt cttttaaatt ttttattggt tgtcgacttg aacggagtga
     1141 caatatatat atatatatat ttaataatga catcattatc tgtaaatctg attcttaatg
     1201 ctattctagt tatgtaagag tggtccccta ggatattgaa gtatccctat cagtgataga
     1261 gaagtcccta tcagtgatag agaatcccta tcagtgatag agatgcgtcc tcgtcttcac
     1321 cggtcgcgtt cctgaaacgc agatgtgcct tccctatcag tgatagagaa acaataaaga
     1381 ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac
     1441 aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga ttagtttttt
     1501 agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat
     1561 ataaatggaa aagctgcaga tctaatcact tcaacttcgc tacaagtagt cactatttgt
     1621 agcaactgta aacgaacaca tcaaagaata agattacatt ctatatctaa gactaaattt
     1681 taaatgtccg ctaaatcgtt tgaagtcaca gatccagtca attcaagtct caaaggcttc
     1741 gcgctagcta acccctccat tacgctggtc cctgaagaaa aaattctctt cagaaagacc
     1801 gattccgaca agatcgcatt aatttctggt ggtggtagtg gacatgaacc tacacacgcc
     1861 ggtttcattg gtaagggtat gttgagtggc gccgtggttg gcgaaatttt tgcatcccct
     1921 tcaacaaaac agattttaaa tgcaatggat ccccgtttag tcaatgaaaa tgcgtctggc
     1981 gttttattga ttgtgaagaa ctacacaggt gatgttttgc attttggtct gtccgctgag
     2041 agagcaagag ccgaattctg tacaaatagg gccgcatcat gtaattagtt atgtcacgct
     2101 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     2161 aagtctaggt ccctatttat tttcttatag ttatgttagt attaagaacg ttatttatat
     2221 ttcaaatttt tctttttttt ctgtacggac gcgtgtacgc atctagctag agcggccgcc
     2281 accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     2341 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2401 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2461 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2821 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2881 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2941 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     3001 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     3061 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     3121 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     3181 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     3241 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     3301 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     3361 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     3421 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     3481 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     3541 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     3601 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     3661 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     3721 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     3781 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     3841 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     3901 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     3961 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     4021 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     4081 gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4141 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4201 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4261 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4321 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4501 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4561 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4621 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4681 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4741 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4801 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4861 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4921 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4981 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5041 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5101 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5161 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5221 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5281 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5341 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5401 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5641 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5881 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5941 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6121 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6181 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6241 gccgaacgac cgagcgcagc gagtcagtg
//
