LOCUS       sequence-241011-        5461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the marked mRNA under the control of insertional 
            promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5461)
  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 5461)
  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_243
FEATURES             Location/Qualifiers
     source          1..5461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..26
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10..26
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        47..65
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     93..109
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     94..110
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    619..637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    698..716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1397..1416)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1398..1416)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1423..1440)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1423..1439)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1432..1454)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1580..2035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1667..1686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1877..1898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2166..2969)
                     /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(2970..3185)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3209..3228)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3328..3350
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3388..3406)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3474..3578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3579..3647,3648..4439)
                     /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(3797..3816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4610..5198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5099..5118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5352..5369
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt aaccctcact
       61 aaagggaaca aaagctggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga
      121 tatcgaattc ctgcagcccg ggggatccac tagtcgcagg ttcatttatc tatgggactt
      181 ctggtaagga actgaaggcg aaggaggaac aattgaagga gaaagccaag atgagagaaa
      241 agttatggat ccaagagctg gagagaaggg aggaagaaac ggaggcaagg agaaaaagag
      301 ccgaattggc aagaatgaag acccttgaga acgaagagga aatcaagaac ttagaaaagg
      361 aactaagcga cctggaaaat aagcttggaa agaagtaatt tggctcttcg atttgctgta
      421 aataaattat gatgagagac aaaagaatga acagctaatt tcgactgtaa tgaagaaatg
      481 taataaaaaa attcatgatg ctatgtccag tattcaagaa aaataattac aaaataacag
      541 cgtacatatg aatattcttt gtttttatct tgtataacta tcacttaaaa catatttata
      601 tcgataggta aatgctattc cctatcagtg atagagaatc aatcgctcta tttacttgat
      661 ttttaatatg tgggtttttt tttgcttgac acatacgtcc ctatcagtga tagagataac
      721 ggtaattgcc agctaacgat cggtacaatt ttcagaaaaa ttgaaacaga gagaatccac
      781 caaatattca attgcaaggt gacgtacttc agataaacac tttattgtac catcttaagc
      841 ggctatataa agtgtcatat acacccttca ccaaatacgc aacatgtctg aatatctagc
      901 tcagacgccc tgcaaattta ctatatggtc aagcgaaata gacctaatca gaacaaacgt
      961 atgtttgatt tatcttactg tggccagatc ggcctttcag tacttctaag gttttatact
     1021 aacttctttt attgatcgtt gtaaactacg gtaacaatta tgtatcaaca gctgttggtg
     1081 aacgctcacc ctttaagcac ggttgggcga ctacttcaat atatccatta tcaaatatac
     1141 aagcaattaa gagctatcta tcaaccagag gaacagtgca cgaactcgga aatacctcac
     1201 acacccttaa atagtatcaa cacttacttc ctaagttacg aaggcagaga gttatctgca
     1261 gatctaacgt gtttattaaa ggacatcacg tcatctagcc acccagattc gaaccatttc
     1321 ataagactac aactggaaaa gcgtacttct ccttccggtt cagcagcggc cgccaccgcg
     1381 gtggagctcc aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
     1441 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     1501 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1561 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1621 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1681 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1741 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1801 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1861 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1921 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1981 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     2041 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga
     2101 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     2161 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     2221 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     2281 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     2341 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     2401 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     2461 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     2521 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     2581 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     2641 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     2701 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     2761 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     2821 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     2881 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     2941 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     3001 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     3061 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     3121 gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     3181 ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3241 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3301 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3361 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3421 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3481 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3541 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3601 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3661 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3721 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3781 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3841 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3901 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3961 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4021 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4081 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4141 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4201 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4261 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4321 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4381 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4441 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4501 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4561 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4621 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4681 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4741 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4801 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4861 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4921 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4981 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5041 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5101 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5161 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5221 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5281 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5341 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5401 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgaa
     5461 c
//
