LOCUS       V015259                 6286 bp    DNA     circular SYN 17-NOV-2020
DEFINITION  Exported.
ACCESSION   V015259
VERSION     V015259
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6286)
  AUTHORS   Frei T, Cella F, Tedeschi F, Gutierrez J, Stan GB, Khammash M,
            Siciliano V.
  TITLE     Characterization and mitigation of gene expression burden in
            mammalian cells
  JOURNAL   Nat Commun 11 (1), 4641 (2020)
   PUBMED   32934213
REFERENCE   2  (bases 1 to 6286)
  AUTHORS   Frei T, Cella F, Tedeschi F, Gutierrez J, Stan G-B., Khammash M,
            Siciliano V.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2020) Synthetic and Systems Biology for
            Biomedicine, Italian Institute of Technology-IIT, Largo Barsanti e
            Martteucci 53, Naples, Campania 80125, Italy
REFERENCE   3  (bases 1 to 6286)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "4641"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-AUG-2020) Synthetic and Systems Biology for Biomedicine, Italian
            Institute of Technology-IIT, Largo Barsanti e Martteucci 53, Naples,
            Campania 80125, Italy"
FEATURES             Location/Qualifiers
     source          1..6286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..266
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     polyA_signal    283..507
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        529..1710
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1741..2301
                     /gene="sigW"
                     /label="ECF RNA polymerase sigma factor SigW"
                     /note="ECF RNA polymerase sigma factor SigW from Bacillus
                     subtilis (strain 168). Accession#: Q45585"
     promoter        complement(2341..2359)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2517..2638
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     regulatory      2728..3017
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(3096..3684)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3858..4715)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4716..4820)
                     /label="AmpR promoter"
     promoter        4890..5206
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5303..5899
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     regulatory      6132..6213
                     /label="SV40 pA"
                     /note="SV40 pA"
                     /regulatory_class="polyA_signal_sequence"
ORIGIN
        1 gtcttctcca tcggatcctg atcaggggag ctcacgggga cagccccccc ccaaagcccc
       61 cagggatgta attacgtccc tcccccgcta gggggcagca gcgaccgccc ggggctccgc
      121 tccggtccgg cgctcccccc gcatcccgag ccggcagcgt gcggggacag cccgggcacg
      181 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
      241 acacctgggg gatacgggga aaaccgcggc atatggcctc gactgtgcct tctagttgcc
      301 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
      361 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
      421 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
      481 atgctgggga tgcggtgggc tctatggctt ctgagggcta gcatcgatgc tccggtgccc
      541 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      601 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      661 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      721 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
      781 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacgcccc
      841 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      901 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      961 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     1021 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     1081 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     1141 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     1201 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     1261 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1321 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1381 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1441 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1501 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1561 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1621 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1681 agtttttttc ttccatttca ggtgtcgtga cataaactgc cctagattaa ttaagccacc
     1741 atggagatga tgattaagaa gcgcatcaag caggtgaaga agggcgacca ggacgccttc
     1801 gccgacatcg tggacatcta caaggacaag atttaccagc tgtgctaccg gatgctgggc
     1861 aacgtgcacg aggccgagga catcgcccag gaggccttca tccgcgccta cgtgaacatc
     1921 gacagcttcg acatcaaccg caagttcagc acctggctgt accgcatcgc caccaacctg
     1981 accatcgacc gcatccgcaa gaagaagccc gactactacc tggacgccga ggtggccggc
     2041 accgagggcc tgaccatgta tagccagatc gtggccgacg gcgtgctgcc cgaggacgcc
     2101 gtggtgagcc tggagctgag caacaccatc cagcagaaga tcctgaagct gcccgacaag
     2161 taccgcaccg tgatcgtgct gaagtacatc gacgagctga gcctgatcga gatcggcgag
     2221 atcctgaaca tccccgtggg caccgtgaag acccgcatcc accgcggccg ggaggccctg
     2281 cgcaagcagc tgcgcgacct gtaaactagt atgtacaagt agtaatctag agggccctat
     2341 tctatagtgt cacctaaatg ctagagctcg ctgatcagtc cgtcgacgga tccaccggat
     2401 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2461 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2521 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2581 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     2641 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2701 tcatttttta accaataggc cgaaatcggg gagctcacgg ggacagcccc cccccaaagc
     2761 ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgaccg cccggggctc
     2821 cgctccggtc cggcgctccc cccgcatccc gagccggcag cgtgcgggga cagcccgggc
     2881 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     2941 cagacacctg ggggatacgg ggaaaatgat caagatcttc tacactggga cgcaaggatt
     3001 atcctcgagg cgcgcgaaga atcaggggat aacgcagtgt gagcaaaagg ccagcaaaag
     3061 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3121 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3181 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3241 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3301 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3361 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3421 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3481 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3541 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3601 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3661 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3721 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3781 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3841 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3901 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3961 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4021 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4081 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4141 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4201 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4261 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4321 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4381 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4441 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     4501 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4561 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4621 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4681 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4741 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4801 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctggagg ttttcaccgt
     4861 catcaccgaa acgcgcgaga cgaaagattg gtgtggaaag tccccaggct ccccagcagg
     4921 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4981 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5041 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5101 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5161 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tagcttgcat
     5221 gcctgcaggt cggccgccac gaccggtgcc gccaccatcc cctgacccac gcccctgacc
     5281 cctcacaagg agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     5341 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     5401 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     5461 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5521 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5581 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5641 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5701 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5761 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5821 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5881 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     5941 cgcacgaccc catggctccg accgaagcca cccggggcgg ccccgccgac cccgcacccg
     6001 cccccgaggc ccaccgactc tagaggatca taatcagcca taccacattt gtagaggttt
     6061 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     6121 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6181 caaatttcac aaataaagca tttttttcac tgcaatctcg tgatacgcct atttttatag
     6241 gttaatgtca tggatgccgc atagttaagc cagtatctgc tccctg
//
