LOCUS       sequence-240991-        6301 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 6301)
  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 6301)
  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_228
FEATURES             Location/Qualifiers
     source          1..6301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    74..95
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(110..115,116..133,134..140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    148..164
                     /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     153..175
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     172..188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     172..188
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        209..227
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1195..1213
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1216..1234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1236..1254
                     /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"
     primer_bind     complement(2076..2094)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2286..2305)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2287..2305)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2312..2329)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2312..2328)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2321..2343)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2469..2924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2556..2575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2766..2787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3055..3858)
                     /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(3859..4074)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4098..4117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4217..4239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4277..4295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4363..4467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4468..4536,4537..5328)
                     /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(4686..4705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5499..6087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5988..6007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6241..6258
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
       61 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      121 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      181 gctatgacca tgattacgcc aagctcgaaa ttaaccctca ctaaagggaa caaaagctgg
      241 taccatcata gagaaatcat atgaaaaatc aaatacaact ctgggcttgg aggaagtcat
      301 tataagatcc ggttacatgg gtgtttgtat tgataacatt ccctcccaat atagctctta
      361 caataatatg actacattct ccaattcaat ttgctatgca agaaagaatt taagctcggt
      421 tcccttatac agagacttgg aaattcaact ctcaaatatt gcatcttcca gctccaaaac
      481 ccaatcaagc gttgtcttga acattttgaa gttagcccaa ttaacttcgg ttaatgttat
      541 acatccatat gtcttgatgg caactgtaat cctaacaatc ctaatgtttc tatttattct
      601 gtatgtaact gtccctaagt taccattcaa actagcggtt aataaatttc tactgttgct
      661 aagttcaact atagttttga catggggtat tggtgcgatg tggactcatg tagggataaa
      721 tgcaagttac agattggttc catcatcaag tatgaatata atcactgtca agaagggcaa
      781 gaaggcagca gtaatggctt ggtttagctt tgcattcctc cttttagaca gtgtggtttt
      841 atggttaata tttttaaggg ataggaaaag cttgaaagat gaaattgata atgttccatg
      901 tgcccaaaat aggtacaata actactcttc ggattcatct acattgcatt ccaaagttta
      961 gctcgagggt cagattcatt atctacaaag actgtaaaaa gaaagtggaa tattcattca
     1021 tatcatattt tttctattaa ctgcctggtt tcttttaaat tttttattgg ttgtcgactt
     1081 gaacggagtg acaatatata tatatatata tttaataatg acatcattat ctgtaaatct
     1141 gattcttaat gctattctag ttatgtaaga gtggtcccct aggatattga agtatcccta
     1201 tcagtgatag agaagtccct atcagtgata gagaatccct atcagtgata gagatgcgtc
     1261 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc ttccctatca gtgatagaga
     1321 aacaataaag attctacaat actagctttt atggttatga agaggaaaaa ttggcagtaa
     1381 cctggcccca caaaccttca aattaacgaa tcaaattaac aaccatagga tgataatgcg
     1441 attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta
     1501 ttaacagata tataaatgga aaagctgcag atcttgcgag aaatcgtatt tagtagtgta
     1561 ttcagttgat aattagtact aattttttat tgcattttgc cttttttagc tttttgtgtt
     1621 acttctagta tccgtaaaaa caatcaaata aatagcactg taaaacatgc ctccaaagaa
     1681 gcaagtcgaa gaaaaaaagg tcttattggg tcgtccagga aacaatttaa aagccggtat
     1741 tgtcggtttg gccaatgttg gtaagtctac ctttttccaa gccatcacta gatgtccatt
     1801 gggtaaccca gctaactatc cattcgctac cattgatcca gaagaagccc gtgttattgt
     1861 cccatctcca agatttgata agttgtgtga aatctacaag aagacagctt cggaagttcc
     1921 agctcatttg gatccgaccg tttacgatat tgctggtttg actaagggtg cctctgctgg
     1981 tgaaggtttg ggtaatgctt tcttgtctca catcagatca gtcgattcta tctacgaatt
     2041 ctgtacaaat agggccgcat catgtaatta gttatgtcac gcttacattc acgccctccc
     2101 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     2161 tattttctta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     2221 tttctgtacg gacgcgtgta cgcatctagc tagagcggcc gccaccgcgg tggagctcca
     2281 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     2341 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2401 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2461 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2521 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2581 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2641 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2701 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2761 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2821 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2881 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2941 ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat
     3001 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     3061 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     3121 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     3181 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     3241 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     3301 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     3361 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     3421 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     3481 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     3541 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     3601 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     3661 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     3721 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     3781 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     3841 atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg
     3901 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     3961 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     4021 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg
     4081 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4141 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4201 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4261 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4321 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4381 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4441 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4501 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4561 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4621 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4681 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4741 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4801 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4861 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4921 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4981 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5041 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5101 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5161 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5221 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5281 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5341 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5401 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5461 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5521 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5581 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5641 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5701 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5761 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5821 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5881 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5941 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6001 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6061 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6121 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6181 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6241 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6301 c
//
