LOCUS       sequence-241265-        6333 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 6333)
  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 6333)
  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: pSM002
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    953..971
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    974..992
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    994..1012
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1060..1078
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1866..1884)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2076..2095)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2077..2095)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2102..2119)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2102..2118)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2111..2133)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2259..2714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2346..2365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2556..2577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2845..3648)
                     /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(3649..3864)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3888..3907)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4007..4029
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4067..4085)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4153..4257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4258..4326,4327..5118)
                     /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(4476..4495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5289..5877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5778..5797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6031..6048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6165..6186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6201..6206,6207..6224,6225..6231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6239..6255
                     /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     6244..6266
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6263..6279
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6263..6279
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6300..6318
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ccatcataga gaaatcatat gaaaaatcaa atacaactct gggcttggag gaagtcatta
       61 taagatccgg ttacatgggt gtttgtattg ataacattcc ctcccaatat agctcttaca
      121 ataatatgac tacattctcc aattcaattt gctatgcaag aaagaattta agctcggttc
      181 ccttatacag agacttggaa attcaactct caaatattgc atcttccagc tccaaaaccc
      241 aatcaagcgt tgtcttgaac attttgaagt tagcccaatt aacttcggtt aatgttatac
      301 atccatatgt cttgatggca actgtaatcc taacaatcct aatgtttcta tttattctgt
      361 atgtaactgt ccctaagtta ccattcaaac tagcggttaa taaatttcta ctgttgctaa
      421 gttcaactat agttttgaca tggggtattg gtgcgatgtg gactcatgta gggataaatg
      481 caagttacag attggttcca tcatcaagta tgaatataat cactgtcaag aagggcaaga
      541 aggcagcagt aatggcttgg tttagctttg cattcctcct tttagacagt gtggttttat
      601 ggttaatatt tttaagggat aggaaaagct tgaaagatga aattgataat gttccatgtg
      661 cccaaaatag gtacaataac tactcttcgg attcatctac attgcattcc aaagtttagc
      721 tcgagggtca gattcattat ctacaaagac tgtaaaaaga aagtggaata ttcattcata
      781 tcatattttt tctattaact gcctggtttc ttttaaattt tttattggtt gtcgacttga
      841 acggagtgac aatatatata tatatatatt taataatgac atcattatct gtaaatctga
      901 ttcttaatgc tattctagtt atgtaagagt ggtcccctag gatattgaag tatccctatc
      961 agtgatagag aagtccctat cagtgataga gaatccctat cagtgataga gatgcgtcct
     1021 cgtcttcacc ggtcgcgttc ctgaaacgca gatgtgcctt ccctatcagt gatagagaaa
     1081 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
     1141 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
     1201 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
     1261 aacagatata taaatggaaa agctgcagat ctatagcatc gctaacatct aaaaaagtta
     1321 tattttcatt gccaattata tttttgagtg ggttatactt gagcattctt caataaaaaa
     1381 gtgttttaaa gaaaaagaag aaagaaattg aagcacgaat aaagttagaa ttaaattcta
     1441 atatgatgca taaaccatgt ttgaagtccc tataacttta actaatagga agtttgctca
     1501 aagaagaaaa ttgaaatacc aatacataaa ttacatttct agaagatttg acagaatttc
     1561 caagaaatct actaccacag atagtttgcc aacgcccgaa aattctgcag ctgaaaataa
     1621 tgacgaagaa gaaggtcaaa acagtgaagc cggcacatat aggaggtcag tactacagca
     1681 gaagaaaaga agaagagaaa ggcattggag gagtgtggtc aattgcggtg aaatttactc
     1741 tactacggaa agcgaaacag actctcaaga ggaggaaact gaggaaggtg gcgaacatga
     1801 caccgggata gacaaggaag atagtgaatt ctgtacaaat agggccgcat catgtaatta
     1861 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     1921 gttagacaac ctgaagtcta ggtccctatt tattttctta tagttatgtt agtattaaga
     1981 acgttattta tatttcaaat ttttcttttt tttctgtacg gacgcgtgta cgcatctagc
     2041 tagagcggcc gccaccgcgg tggagctcca attcgcccta tagtgagtcg tattacaatt
     2101 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2161 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2221 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
     2281 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2341 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2401 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2461 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2521 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2581 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2641 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2701 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2761 cgcatagggt aataactgat ataattaaat tgaagctcta atttgtgagt ttagtataca
     2821 tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc
     2881 ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag
     2941 tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata
     3001 ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca
     3061 atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc
     3121 tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc
     3181 cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc
     3241 cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc
     3301 tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat
     3361 gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag
     3421 cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa
     3481 acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc
     3541 caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac
     3601 aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg
     3661 tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
     3721 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     3781 cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga
     3841 ataaaaaaaa aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct
     3901 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3961 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4021 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4081 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4141 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4201 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4261 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4321 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4381 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4441 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4501 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4561 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4621 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4681 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4741 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4801 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4861 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4921 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4981 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5041 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5101 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5161 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5221 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5281 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5341 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5401 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5461 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5521 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5581 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5641 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5701 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5761 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5821 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5881 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5941 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6001 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6061 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6121 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6181 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6241 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgaa
     6301 attaaccctc actaaaggga acaaaagctg gta
//
