LOCUS       sequence-241259-        9095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the repressor to control the insertional promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9095)
  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 9095)
  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_238
FEATURES             Location/Qualifiers
     source          1..9095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..82
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    199..220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(235..240,241..258,259..265)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    273..289
                     /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     278..300
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     297..313
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     297..313
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        334..352
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1094..1714
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MPRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1167..1186)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(5122..5141)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5123..5141)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5148..5165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5148..5164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5157..5179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5305..5760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5392..5411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5602..5623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6020..6682)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(6683..6870)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(7017..7036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7136..7158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7196..7214)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7282..7386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7387..7455,7456..8247)
                     /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(7605..7624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8418..9006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8907..8926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
       61 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      121 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      181 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      241 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      301 aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa
      361 gctggtaccg ggccccccct cgaggtcgac attacgatgg cttcttatct cacttcaata
      421 gtactttcca ccggttatac ttccggcttt tccctattaa tacaagctac aatttcaatg
      481 ggtggcaaat aatgtgtaga atagaaaata agccgacagg gtaataaaga aaatttttag
      541 aaaaaaaagg ttagatggct tatttaagtt acaggctagc gaaaaaagga acttcagggc
      601 aagtaaagtg tttgattggg cactagcatg gcttataaag gcgagcaatt gtcgaaacta
      661 attaatgttg tacggactat tgctgtcttc tcgtggtaaa tgcgtgttcc aggtcgaata
      721 ctacttgcac acaggcgagc ggggccccat aaaagtgttg ccgatttgtt aagttgtctt
      781 ttcggttttt ctactctgtt attccttact tcccttttta agaactcttt ttatccttca
      841 tttaggatct tgcacgtttc cgcctcatca cttgaattaa aacatgtctc tgtcagtaaa
      901 ccttggcgtt tctattgttc ttcatagttc aacttttatt attacccgcc ctgcgcgttt
      961 acatttttcc agcaacagcc agcgaaaaat tagaaaatct ggttgttgac acctcaagaa
     1021 caagggcaat tagcctcagc gtcgaatata gatcatatta gaatacctat agctccatca
     1081 aaagaaatac aaaatgccta gattagataa aagtaaagtg attaacagcg cattagagct
     1141 gcttaatggg gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt
     1201 agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttacc
     1261 cattgagatg ttagataggc accatactca cttttgccct ttagaagggg aaagctggca
     1321 agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg
     1381 agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca
     1441 attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc
     1501 tgtggggcac tttactttag gttgcgtatt ggaagaacaa gagcatcaag tcgctaaaga
     1561 agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt
     1621 atttgatcgc caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg
     1681 attagaaaaa caacttaaat gtgaaagtgg gtccgcgtcc gcgggcgcat ctgagaacac
     1741 cacagcccct tctgataaca taaccaatga acagagactt ccgtctggac ccaaagatga
     1801 cgtagatacg ttagcactta ccagtgctca aaatcaagct aatagtttga ggaagctaga
     1861 tacagatgcg aacgctaaag ctttgccatc catcacagac ataccagtca gtgatgattt
     1921 tgatataaaa cgacaagttg gctctggttt tggttcaaat ccgcttcata taaaagattc
     1981 cgaggccttt ccacattcat ctattgaagc cctaaaagag ggtatggaca aagtcacaaa
     2041 acaatgcaac gatttgaaga cggctttgct ttccaaagac acttccctca ctgattctgt
     2101 gcaggatttg tttaactcat taaaagtttt atcacacaac caatcagttt tggaaaataa
     2161 gctggatgat gtcatgaaga atcaagtaaa tactgacata ttggtcaata atttgaatga
     2221 acgattgaac aaattatcga caatgttgca aaatacttcg aaagtaaatc actctaacct
     2281 tctcatagaa aattcatcca acaataccag ttcacagcat aatacctctt cttctcgaag
     2341 gggacctggc aggccaagaa aagatgcctc gacctccaca atgaataagt tagtgtcaaa
     2401 cgcagcctcg gtcaatctca aaagtgcatc caatcagggt gctcctttct caccagtaaa
     2461 tattacttta ccgactgccg tagtacaaac gtctaaatcc aaaaggtatt ttgtggaacc
     2521 atcaacgaaa caagagtcgc ttttattatc tgccccttca tcatcaagag atgatgctga
     2581 tatgtccttg acttccgtac cgcaaagaac gaacaatgaa aatggtaaag agcgaccatc
     2641 cactgctaat tctagctcca tcacaccaac acctgttacg ccgaacaact taattcaaat
     2701 caaaagaaaa agaggcaggc ccccaaaaaa gagaacagtg gaaacaatga tatccaattc
     2761 cacggatacg atagataagt cagatgcatc taatcggatt aaaaacgaga ttccaataaa
     2821 ttccttgctt ccatcttcca aatttcatca aataccatca tctccatcca atcctgtgtc
     2881 acaaccggct ccggttcgga cttcaaggtc agccacacaa gaaatagaca ttaaaagttt
     2941 agaactggcc tctctaattt caactaatgg tgatccaaac gcagaggatt cgaatactac
     3001 tgatactgtt cataataacg tggaaggaaa ggtaaatgtt gaagaaaata aaaccgagaa
     3061 ggagaaaata ataaccatca aatcatccag cgaaaatagt ggtaataata cgaccaataa
     3121 taataatact gacaacgtca ttaaattttc agctaattca gatatcaata gtgatattcg
     3181 ccgattaatg gttaacgatc agttttcatt aagttatgac gccagcggta atattacggt
     3241 caaattacca cccgtttcct ctccagcagc ggcaacagct gccgctgctg cggcggttac
     3301 gtcagagatg aatagacaac aaagagaatt agataaaaga cgtgattcaa gggagaaaat
     3361 gcttgttaat atgaaatata acgatcgcga taaagcaaag tcatttatgg agtctaataa
     3421 gaaactctta aaagcaatga aagaagaaga gaggaggaaa agaatgactt cgataataca
     3481 tgataatcac ttgaacttaa atttgaatga gatctccacc cgttcaaaga taaaaagtgc
     3541 agaaaaacca acaactaaag gttcttcaat gtcgccaaaa ccaagatcgg cctccatcag
     3601 cggcatttca gaccaccaac aggaaggata tcaaccatta gagcaagaaa aactcgtcga
     3661 tattgacaat gagggctcaa acgcaaacag cgactctctc aagatgggtc taaccatatc
     3721 cgctgccgat acggttcaca aagttggaat acagtccatg ctaaattctg gggaagaggc
     3781 aattaccaag gagaatgcag aatatgaaag aaagaccccg ggagatgaag aaactaccac
     3841 gttcgttcca ttagaaaact cacaaccttc ggacacgata aggaaaagaa cagcaggcga
     3901 tgacggtgca ttggatcaaa cagagaatac gagcatatca ccaaagaaga gacgcacaga
     3961 agatcataca aaaggtgaag aagatgaggg agaaaggggt gttggtaata gcggaacgct
     4021 ggccaccgtg gagaatgtct caggagacat ctccgcagat ttatccaaag gtacttcatc
     4081 tatccataac gatactgagt ctgctaacga tagtagcaat ggcaacggaa acctcggttt
     4141 gggtacagag tcacgcaata cattgttaac ggcaactccc attgaattaa tctgccggga
     4201 aggcttcttc taccggagag atattccgga cgttcccatt accacaggag cgtacctgga
     4261 atttaaattc aaggcgaaag aagaggaatt gatcaactct agcatcaatg aggaggatta
     4321 tgctgctaaa tcaaagcatg aaaaaatgaa tgcgcatttc ttcaagcctg atattcaaga
     4381 agagacggaa cttgcctttg aaattttgag caagacaacg ctgacagaga aatacgttaa
     4441 cagtttggag tacttcttga tggaattcag gtgggagaat aaactagttg gtctaggctt
     4501 aaaacttcgg gaatccaaaa gaacctggca aaggagaaag gcgttatttg cccttttcga
     4561 attttggaga gaccaatcaa gagacaaaag aagattccac aactacacca tcctgcatgc
     4621 ggtgaaagag atggaaaact acagaatatt tattaatcga tccgtttcat ggttctacaa
     4681 ccatattacc ctgctaaaaa tgatcctcta cgacctctgc gacaatgtga ccactcaatg
     4741 gagagagtgg atgtttcccc ataacgaaac actgccggca ttgggtcagg acggcattaa
     4801 cgaagacaat ctgaatgaaa ccatcgacaa catgttaatt tttgacttcc ttgatgacgg
     4861 ttcagaaaac aaccaggtca aatattccag aatcataccg ccagatatcc gttaacaatt
     4921 caaaaatatg gactgcattt gaactagtgt ttctaatctc tgcttttgtg cgcgtatgtt
     4981 tatgtatgta cctctctctc tatttctatt tttaaaccac cctctcaata aaataaaaat
     5041 aataaagtat ttttaaggaa aagacgtgtt taagcactga ctttatctac gcggccgcca
     5101 ccgcggtgga gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
     5161 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     5221 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5281 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5341 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5401 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5461 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5521 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5581 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5641 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5701 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5761 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagatccgtc
     5821 gagttcaaga gaaaaaaaaa gaaaaagcaa aaagaaaaaa ggaaagcgcg cctcgttcag
     5881 aatgacacgt atagaatgat gcattacctt gtcatcttca gtatcatact gttcgtatac
     5941 atacttactg acattcatag gtatacatat atacacatgt atatatatcg tatgctgcag
     6001 ctttaaataa tcggtgtcac tacataagaa cacctttggt ggagggaaca tcgttggtac
     6061 cattgggcga ggtggcttct cttatggcaa ccgcaagagc cttgaacgca ctctcactac
     6121 ggtgatgatc attcttgcct cgcagacaat caacgtggag ggtaattctg ctagcctctg
     6181 caaagctttc aagaaaatgc gggatcatct cgcaagagag atctcctact ttctcccttt
     6241 gcaaaccaag ttcgacaact gcgtacggcc tgttcgaaag atctaccacc gctctggaaa
     6301 gtgcctcatc caaaggcgca aatcctgatc caaacctttt tactccacgc acggccccta
     6361 gggcctcttt aaaagcttga ccgagagcaa tcccgcagtc ttcagtggtg tgatggtcgt
     6421 ctatgtgtaa gtcaccaatg cactcaacga ttagcgacca gccggaatgc ttggccagag
     6481 catgtatcat atggtccaga aaccctatac ctgtgtggac gttaatcact tgcgattgtg
     6541 tggcctgttc tgctactgct tctgcctctt tttctgggaa gatcgagtgc tctatcgcta
     6601 ggggaccacc ctttaaagag atcgcaatct gaatcttggt ttcatttgta atacgcttta
     6661 ctagggcttt ctgctctgtc atctttgcct tcgtttatct tgcctgctca ttttttagta
     6721 tattcttcga agaaatcaca ttactttata taatgtataa ttcattatgt gataatgcca
     6781 atcgctaaga aaaaaaaaga gtcatccgct aggtggaaaa aaaaaaatga aaatcattac
     6841 cgaggcataa aaaaatatag agtgtactag aggaggccaa gagtaataga aaaagaaaat
     6901 tgcgggaaag gactgtgtta tgacttccct gactaatgcc gtgttcaaac gatacctggc
     6961 agtgactcct agcgctcacc aagctcttaa aacgggaatt tatggtgcac tctcagtaca
     7021 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     7081 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7141 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     7201 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7381 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7441 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7501 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7561 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7621 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7681 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7741 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7801 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7861 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7921 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7981 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     8041 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8101 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8161 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8221 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8281 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8341 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     9001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     9061 cacatgttct ttcctgcgtt atcccctgat tctgt
//
