LOCUS       V015442                 6570 bp    DNA     circular SYN 30-JUN-2020
DEFINITION  Exported.
ACCESSION   V015442
VERSION     V015442
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6570)
  AUTHORS   Scheller L, Schmollack M, Bertschi A, Mansouri M, Saxena P,
            Fussenegger M.
  TITLE     Phosphoregulated orthogonal signal transduction in mammalian cells
  JOURNAL   Nat Commun 11 (1), 3085 (2020)
   PUBMED   32555187
REFERENCE   2  (bases 1 to 6570)
  AUTHORS   Scheller L.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment
            AI), Station 19, Lausanne 1015, Switzerland
REFERENCE   3  (bases 1 to 6570)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "3085"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment AI), Station
            19, Lausanne 1015, Switzerland"
FEATURES             Location/Qualifiers
     source          1..6570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..419
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        420..623
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        668..686
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    715..720
                     /label="Kozak+ATG"
                     /note="Kozak+ATG"
     CDS             739..1455
                     /gene="dcuR"
                     /label="Transcriptional regulatory protein DcuR"
                     /note="Transcriptional regulatory protein DcuR from
                     Escherichia coli (strain K12). Accession#: P0AD01"
     CDS             1492..2109
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     polyA_signal    2170..2394
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2440..2868
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2882..3211
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3278..4069
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4246..4379
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4416..4432)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4440..4456)
                     /label="lac operator"
                     /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)."
     promoter        complement(4464..4494)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4509..4530)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4818..5403)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5577..6434)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6435..6539)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctcca cgcgtggtac cctcgagttg acattgatta ttgagtagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      601 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      661 tcgaaattaa tacgactcac tatagggaga cccaagctgt tcgaagcgga attcaccatg
      721 actagtggtg gttctggtat gatcaatgta ttaattatcg atgacgacgc aatggtcgcg
      781 gagctgaatc gccgatacgt agcacaaatc ccaggctttc aatgctgtgg aacagcctcg
      841 acgctggaga aagccaaaga gattatcttc aatagcgata cgcctatcga cctgatattg
      901 ctcgatatct atatgcaaaa agagaacggg ctcgatttac tgcctgtcct gcataacgcg
      961 cgttgcaaaa gtgatgtgat tgtcatctcc tccgcagccg atgcggcaac cattaaagat
     1021 tcgctgcatt acggtgtcgt ggattacctg atcaaaccct tccaggcttc ccgctttgaa
     1081 gaggcgctca ccggctggcg gcaaaagaaa atggcgctgg aaaaacatca gtattatgac
     1141 caggccgagc ttgatcagct aatccacggc agcagctcca acgaacagga tcctcgtcgc
     1201 ttgccaaaag gcttaacgcc gcagacgtta cgcacgctgt gccagtggat tgacgcgcat
     1261 caggactatg agttttcaac cgacgagcta gccaacgagg ttaacatttc gcgtgtttcc
     1321 tgccgtaaat acctcatctg gttggtcaac tgccacatct tgttcaccag tatccattat
     1381 ggcgtcacgg ggcgtccagt ttaccgttac cgtattcagg cagaacacta ctcactactg
     1441 aaacaatatt gccaagctag cggatccacc ggttctagtg gtggttctgg ttccagatta
     1501 gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg aatcgaaggt
     1561 ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt gtattggcat
     1621 gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga taggcaccat
     1681 actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa taacgctaaa
     1741 agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt aggtacacgg
     1801 cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg ccaacaaggt
     1861 ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac tttaggttgc
     1921 gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc tactactgat
     1981 agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg tgcagagcca
     2041 gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact taaatgtgaa
     2101 agtgggtccg ctagcggatc caccggtgtc tagaaagctt tgaggccggc ctgcagggat
     2161 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2221 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2281 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2341 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2401 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2461 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2521 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2581 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2641 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2701 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2761 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2821 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2881 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3001 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3061 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3121 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3181 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3241 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3301 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3361 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3421 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3481 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3541 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3601 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3661 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3721 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3781 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3841 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3901 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3961 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4021 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4081 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4141 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4201 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4261 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4321 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4381 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4441 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4501 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4561 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4621 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4681 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4741 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5221 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5341 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5641 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5701 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5761 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5821 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5881 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5941 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6001 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6061 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6181 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6541 acatttcccc gaaaagtgcc acctgacgtc
//
