pGLM103(P_EF1a-tTA--Cer) vector (V016453)

LOCUS       62056_11770        7268 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pGLM103(P_EF1a-tTA--Cer), complete sequence.
ACCESSION   MT891338
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7268)
  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 7268)
  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 7268)
  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..7268
                     /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..2484
                     /label=tTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
     CDS             2491..3207
                     /label=mCerulean
                     /note="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
     misc_binding    3221..3242
                     /label=miR-FF5 binding site
                     /bound_moiety="miR-FF5"
                     /note="mir FF5 target"
     misc_binding    3243..3264
                     /label=miR-FF5 binding site
                     /bound_moiety="miR-FF5"
                     /note="mir FF5 target"
     misc_binding    3265..3286
                     /label=miR-FF5 binding site
                     /bound_moiety="miR-FF5"
                     /note="mir FF5 target"
     promoter        complement(3323..3341)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3499..3620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3710..3999
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(4078..4666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4840..5697)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5698..5802)
                     /label=AmpR promoter
     promoter        5872..6188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6285..6881
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     regulatory      7114..7195
                     /label=SV40 pA
                     /note="SV40 pA"
                     /regulatory_class="polyA_signal_sequence"