LOCUS       62056_21        4123 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (181)_P_EFS-mRuby3, complete sequence.
ACCESSION   MT891328
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4123)
  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 4123)
  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 4123)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "4641"
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4123
                     /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..740
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             779..1489
                     /label=mRuby3
                     /note="monomeric red fluorescent protein derived from
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
     promoter        complement(1511..1529)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1690..1811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1901..2190
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(2278..2866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3040..3897)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3898..4002)
                     /label=AmpR promoter
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 gcatcgatgg gcagagcgca
      541 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
      601 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      661 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      721 ggtttgccgc cagaacacag cataaactgc cctagattaa ttaagcggcc gcgccaccat
      781 ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg
      841 ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcagac cgtacgaggg
      901 agtgcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat
      961 tcttgccacg tcgttcatgt atggcagccg tacctttatc aagtacccgg ccgacatccc
     1021 tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga
     1081 agatggtgga gtcgtcaccg tcacgcagga caccagcctt gaggatggcg agctcgtcta
     1141 caacgtcaag gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac
     1201 caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc tgagaggata
     1261 cactgacatc gcactgaaag ttgatggtgg tggccatctg cactgcaact tcgtgacaac
     1321 ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtgtccatg ccgttgatca
     1381 ccgcctggaa aggatcgagg agagtgacaa tgaaacctac gtagtgcaaa gagaagtggc
     1441 agttgccaaa tacagcaacc ttggtggtgg catggacgag ctgtacaagt agtaatctag
     1501 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagtc catagtcgac
     1561 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     1621 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1681 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1741 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1801 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1861 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggggagctca cggggacagc
     1921 ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga
     1981 ccgcccgggg ctccgctccg gtccggcgct ccccccgcat cccgagccgg cagcgtgcgg
     2041 ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg cttctcgctg
     2101 ctctttgagc ctgcagacac ctgggggata cggggaaaat gatcaagatc ttctacactg
     2161 ggacgcaagg attatcctcg aggcgcgcga agaatcaggg gataacgcag gaaagaacat
     2221 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2281 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2341 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2401 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2461 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2521 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2581 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2641 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2701 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2761 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3961 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4021 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     4081 atctgctctg atgccgcata gttaagccag tatctgctcc ctg
//
