LOCUS       V016382                 7599 bp    DNA     circular SYN 30-JUN-2020
DEFINITION  Exported.
ACCESSION   V016382
VERSION     V016382
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7599)
  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 7599)
  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 7599)
  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..7599
                     /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             730..750
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             754..774
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             778..798
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             817..1533
                     /gene="dcuR"
                     /label="Transcriptional regulatory protein DcuR"
                     /note="Transcriptional regulatory protein DcuR from
                     Escherichia coli (strain K12). Accession#: P0AD01"
     CDS             1573..1722
                     /label="VP64"
                     /note="tetrameric repeat of the minimal activation domain
                     of herpes simplex virus VP16 (Beerli et al., 1998)"
     CDS             1747..1767
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2197..2553
                     /label="RelA (p65) AD"
                     /note="transcriptional activation domain of human RelA,
                     also known as p65 (O'Shea and Perkins, 2008)"
     CDS             2572..3138
                     /label="Rta AD"
                     /note="transcriptional activation domain from the human
                     herpesvirus 4 (Epstein-Barr virus) replication and
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992;
                     Chavez et al., 2015)"
     polyA_signal    3199..3423
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3469..3897
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3911..4240
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             4307..5098
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5275..5408
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5445..5461)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5469..5485)
                     /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(5493..5523)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5538..5559)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5847..6432)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6606..7463)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7464..7568)
                     /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 actagtgatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa ggtagatcca
      781 aaaaagaaga gaaaggtagc tagtggtggt tctggtatga tcaatgtatt aattatcgat
      841 gacgacgcaa tggtcgcgga gctgaatcgc cgatacgtag cacaaatccc aggctttcaa
      901 tgctgtggaa cagcctcgac gctggagaaa gccaaagaga ttatcttcaa tagcgatacg
      961 cctatcgacc tgatattgct cgatatctat atgcaaaaag agaacgggct cgatttactg
     1021 cctgtcctgc ataacgcgcg ttgcaaaagt gatgtgattg tcatctcctc cgcagccgat
     1081 gcggcaacca ttaaagattc gctgcattac ggtgtcgtgg attacctgat caaacccttc
     1141 caggcttccc gctttgaaga ggcgctcacc ggctggcggc aaaagaaaat ggcgctggaa
     1201 aaacatcagt attatgacca ggccgagctt gatcagctaa tccacggcag cagctccaac
     1261 gaacaggatc ctcgtcgctt gccaaaaggc ttaacgccgc agacgttacg cacgctgtgc
     1321 cagtggattg acgcgcatca ggactatgag ttttcaaccg acgagctagc caacgaggtt
     1381 aacatttcgc gtgtttcctg ccgtaaatac ctcatctggt tggtcaactg ccacatcttg
     1441 ttcaccagta tccattatgg cgtcacgggg cgtccagttt accgttaccg tattcaggca
     1501 gaacactact cactactgaa acaatattgc caagctagcg gatccaccgg ttctagtggt
     1561 tccggacggg ctgacgcatt ggacgatttt gatctggata tgctgggaag tgacgccctc
     1621 gatgattttg accttgacat gcttggttcg gatgcccttg atgactttga cctcgacatg
     1681 ctcggcagtg acgcccttga tgatttcgac ctggacatgc tgattaactc tagaagttcc
     1741 ggatctccga aaaagaaacg caaagttggt agccagtacc tgcccgacac cgacgaccgg
     1801 caccggatcg aggaaaagcg gaagcggacc tacgagacat tcaagagcat catgaagaag
     1861 tcccccttca gcggccccac cgaccctaga cctccaccta gaagaatcgc cgtgcccagc
     1921 agatccagcg ccagcgtgcc aaaacctgcc ccccagcctt accccttcac cagcagcctg
     1981 agcaccatca actacgacga gttccctacc atggtgttcc ccagcggcca gatctctcag
     2041 gcctctgctc tggctccagc ccctcctcag gtgctgcctc aggctcctgc tcctgcacca
     2101 gctccagcca tggtgtctgc actggctcag gcaccagcac ccgtgcctgt gctggctcct
     2161 ggacctccac aggctgtggc tccaccagcc cctaaaccta cacaggccgg cgagggcaca
     2221 ctgtctgaag ctctgctgca gctgcagttc gacgacgagg atctgggagc cctgctggga
     2281 aacagcaccg atcctgccgt gttcaccgac ctggccagcg tggacaacag cgagttccag
     2341 cagctgctga accagggcat ccctgtggcc cctcacacca ccgagcccat gctgatggaa
     2401 taccccgagg ccatcacccg gctcgtgaca ggcgctcaga ggcctcctga tccagctcct
     2461 gcccctctgg gagcaccagg cctgcctaat ggactgctgt ctggcgacga ggacttcagc
     2521 tctatcgccg atatggattt ctcagccttg ctgggctctg gcagcggcag ccgggattcc
     2581 agggaaggga tgtttttgcc gaagcctgag gccggctccg ctattagtga cgtgtttgag
     2641 ggccgcgagg tgtgccagcc aaaacgaatc cggccatttc atcctccagg aagtccatgg
     2701 gccaaccgcc cactccccgc cagcctcgca ccaacaccaa ccggtccagt acatgagcca
     2761 gtcgggtcac tgaccccggc accagtccct cagccactgg atccagcgcc cgcagtgact
     2821 cccgaggcca gtcacctgtt ggaggatccc gatgaagaga cgagccaggc tgtcaaagcc
     2881 cttcgggaga tggccgatac tgtgattccc cagaaggaag aggctgcaat ctgtggccaa
     2941 atggaccttt cccatccgcc cccaaggggc catctggatg agctgacaac cacacttgag
     3001 tccatgaccg aggatctgaa cctggactca cccctgaccc cggaattgaa cgagattctg
     3061 gataccttcc tgaacgacga gtgcctcttg catgccatgc atatcagcac aggactgtcc
     3121 atcttcgaca catctctggc tagcggatcc accggtgtct agaaagcttt gaggccggcc
     3181 tgcagggatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3241 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3301 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3361 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3421 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3481 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3541 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3601 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3661 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3721 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3781 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3841 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3901 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3961 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4021 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4081 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4141 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4201 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4261 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4321 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4381 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4441 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4501 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4561 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4621 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4681 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4741 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4801 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4861 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4921 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4981 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5041 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5101 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5161 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5221 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5281 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5341 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5401 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5461 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5521 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5581 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5641 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5701 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5761 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5821 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5881 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5941 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6001 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6061 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6121 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6181 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6241 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6301 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6361 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6421 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6481 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6541 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6601 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6661 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6721 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6781 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6841 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6901 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6961 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7021 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7081 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7141 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7201 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7261 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7321 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7381 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7441 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7501 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7561 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
