LOCUS       sequence-241264-        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: pSM001
FEATURES             Location/Qualifiers
     source          1..6269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..39
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    156..177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(192..197,198..215,216..222)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     254..270
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     254..270
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        291..309
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1277..1295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1298..1316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1318..1336
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1384..1402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2126..2144)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2336..2355)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2337..2355)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2362..2379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2362..2378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2371..2393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2519..2974
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2606..2625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2816..2837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3105..3908)
                     /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(3909..4124)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4148..4167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4267..4289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4327..4345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4413..4517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4518..4586,4587..5378)
                     /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(4736..4755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5549..6137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6038..6057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
       61 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
      121 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      181 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcga aattaaccct
      301 cactaaaggg aacaaaagct ggtaccatca tagagaaatc atatgaaaaa tcaaatacaa
      361 ctctgggctt ggaggaagtc attataagat ccggttacat gggtgtttgt attgataaca
      421 ttccctccca atatagctct tacaataata tgactacatt ctccaattca atttgctatg
      481 caagaaagaa tttaagctcg gttcccttat acagagactt ggaaattcaa ctctcaaata
      541 ttgcatcttc cagctccaaa acccaatcaa gcgttgtctt gaacattttg aagttagccc
      601 aattaacttc ggttaatgtt atacatccat atgtcttgat ggcaactgta atcctaacaa
      661 tcctaatgtt tctatttatt ctgtatgtaa ctgtccctaa gttaccattc aaactagcgg
      721 ttaataaatt tctactgttg ctaagttcaa ctatagtttt gacatggggt attggtgcga
      781 tgtggactca tgtagggata aatgcaagtt acagattggt tccatcatca agtatgaata
      841 taatcactgt caagaagggc aagaaggcag cagtaatggc ttggtttagc tttgcattcc
      901 tccttttaga cagtgtggtt ttatggttaa tatttttaag ggataggaaa agcttgaaag
      961 atgaaattga taatgttcca tgtgcccaaa ataggtacaa taactactct tcggattcat
     1021 ctacattgca ttccaaagtt tagctcgagg gtcagattca ttatctacaa agactgtaaa
     1081 aagaaagtgg aatattcatt catatcatat tttttctatt aactgcctgg tttcttttaa
     1141 attttttatt ggttgtcgac ttgaacggag tgacaatata tatatatata tatttaataa
     1201 tgacatcatt atctgtaaat ctgattctta atgctattct agttatgtaa gagtggtccc
     1261 ctaggatatt gaagtatccc tatcagtgat agagaagtcc ctatcagtga tagagaatcc
     1321 ctatcagtga tagagatgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg
     1381 ccttccctat cagtgataga gaaacaataa agattctaca atactagctt ttatggttat
     1441 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
     1501 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
     1561 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc agatctatat
     1621 tcaattgcaa ggtgacgtac ttcagataaa cactttattg taccatctta agcggctata
     1681 taaagtgtca tatacaccct tcaccaaata cgcaacatgt ctgaatatct agctcagacg
     1741 ccctgcaaat ttactatatg gtcaagcgaa atagacctaa tcagaacaaa cctgttggtg
     1801 aacgctcacc ctttaagcac ggttgggcga ctacttcaat atatccatta tcaaatatac
     1861 aagcaattaa gagctatcta tcaaccagag gaacagtgca cgaactcgga aatacctcac
     1921 acacccttaa atagtatcaa cacttacttc ctaagttacg aaggcagaga gttatctgca
     1981 tcgataacgt gtttattaaa ggacatcacg tcatctagcc acccagattc gaaccatttc
     2041 ataagactac aactggaaaa gcgtacttct ccttccggtt cagcagaatt ctgtacaaat
     2101 agggccgcat catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     2161 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttctta
     2221 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtacg
     2281 gacgcgtgta cgcatctagc tagagcggcc gccaccgcgg tggagctcca attcgcccta
     2341 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2401 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2461 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     2521 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2581 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2641 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2761 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2821 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2881 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2941 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     3001 atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat tgaagctcta
     3061 atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg
     3121 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
     3181 tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc
     3241 acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca
     3301 ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca
     3361 ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat
     3421 tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt
     3481 tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca
     3541 ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc
     3601 aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac
     3661 ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata
     3721 tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat
     3781 tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata
     3841 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
     3901 ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat
     3961 actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg
     4021 tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga
     4081 aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca
     4141 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4201 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4261 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4321 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4381 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6241 ccgcctttga gtgagctgat accgctcgc
//
