(145)_sp(A)-mRuby3-P_bi vector (V016516)

LOCUS       62056_16        6694 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (145)_sp(A)-mRuby3-P_bi, complete sequence.
ACCESSION   MT891327
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6694)
  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 6694)
  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 6694)
  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..6694
                     /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"
     polyA_signal    complement(525..573)
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     CDS             complement(601..1311)
                     /label=mRuby3
                     /note="monomeric red fluorescent protein derived from
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
     promoter        complement(1345..1732)
                     /label=bidirectional TRE promoter
                     /note="Tet-responsive bidirectional promoter PTight-BI, 
                     consisting of seven tet operator sequences flanked on each 
                     side by the minimal CMV promoter"
     CDS             1769..2713
                     /label=ORF frame 1
                     /note="bright monomeric near-infrared fluorescent protein 
                     engineered from a bacterial phytochrome, with an emission 
                     maximum of 670 nm (Shcherbakova et al., 2016)"
     promoter        complement(2746..2764)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2925..3046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3136..3425
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(3504..4092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4266..5123)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5124..5228)
                     /label=AmpR promoter
     promoter        5298..5614
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5711..6307
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     regulatory      6540..6621
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /regulatory_class="polyA_signal_sequence"