LOCUS       sequence-234494-        5576 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 5576)
  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 5576)
  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..5576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2289..2305
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2290..2306
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2815..2833
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2894..2912
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(3593..3612)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3594..3612)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3619..3636)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3619..3635)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3628..3650)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3776..4231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3863..3882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4073..4094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4362..5165)
                     /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(5166..5381)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5407..5426)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5526..5548
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtaccgg
     2281 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
     2341 atccactagt cgcaggttca tttatctatg ggacttctgg taaggaactg aaggcgaagg
     2401 aggaacaatt gaaggagaaa gccaagatga gagaaaagtt atggatccaa gagctggaga
     2461 gaagggagga agaaacggag gcaaggagaa aaagagccga attggcaaga atgaagaccc
     2521 ttgagaacga agaggaaatc aagaacttag aaaaggaact aagcgacctg gaaaataagc
     2581 ttggaaagaa gtaatttggc tcttcgattt gctgtaaata aattatgatg agagacaaaa
     2641 gaatgaacag ctaatttcga ctgtaatgaa gaaatgtaat aaaaaaattc atgatgctat
     2701 gtccagtatt caagaaaaat aattacaaaa taacagcgta catatgaata ttctttgttt
     2761 ttatcttgta taactatcac ttaaaacata tttatatcga taggtaaatg ctattcccta
     2821 tcagtgatag agaatcaatc gctctattta cttgattttt aatatgtggg tttttttttg
     2881 cttgacacat acgtccctat cagtgataga gataacggta attgccagct aacgatcggt
     2941 acaattttca gaaaaattga aacagagaga atccaccaaa tattcaattg caaggtgacg
     3001 tacttcagat aaacacttta ttgtaccatc ttaagcggct atataaagtg tcatatacac
     3061 ccttcaccaa atacgcaaca tgtctgaata tctagctcag acgccctgca aatttactat
     3121 atggtcaagc gaaatagacc taatcagaac aaacgtatgt ttgatttatc ttactgtggc
     3181 cagatcggcc tttcagtact tctaaggttt tatactaact tcttttattg atcgttgtaa
     3241 actacggtaa caattatgta tcaacagctg ttggtgaacg ctcacccttt aagcacggtt
     3301 gggcgactac ttcaatatat ccattatcaa atatacaagc aattaagagc tatctatcaa
     3361 ccagaggaac agtgcacgaa ctcggaaata cctcacacac ccttaaatag tatcaacact
     3421 tacttcctaa gttacgaagg cagagagtta tctgcagatc taacgtgttt attaaaggac
     3481 atcacgtcat ctagccaccc agattcgaac catttcataa gactacaact ggaaaagcgt
     3541 acttctcctt ccggttcagc agcggccgcc accgcggtgg agctccaatt cgccctatag
     3601 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     3661 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     3721 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     3781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3901 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3961 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4081 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4141 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4201 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     4261 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     4321 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     4381 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     4441 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     4501 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     4561 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     4621 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     4681 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     4741 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     4801 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     4861 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     4921 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     4981 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     5041 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     5101 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     5161 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     5221 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     5281 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     5341 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tatggtgcac
     5401 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5461 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5521 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
