(219)_P_TRE-(dHDV)mCitr vector (V016129)

LOCUS       62056_36        9978 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (219)_P_TRE-(dHDV)mCitr, complete sequence.
ACCESSION   MT891331
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9978)
  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 9978)
  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 9978)
  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..9978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..35
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac left (5') inverted repeat"
     misc_feature    361..385
                     /label=ConS scar
                     /note="ConS scar"
     misc_feature    390..414
                     /label=bidirectional promoter artefact
                     /note="bidirectional promoter artefact"
     promoter        415..729
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     misc_feature    767..919
                     /label=dHDV
                     /note="dHDV"
     CDS             917..1630
                     /label=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     regulatory      1668..1789
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    1891..1915
                     /label=Con1 scar
                     /note="Con1 scar"
     regulatory      1954..2219
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     polyA_signal    2236..2460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2461..3642
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3665..4408
                     /label=tTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
     CDS             4484..5194
                     /label=mRuby3
                     /note="monomeric red fluorescent protein derived from
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
     polyA_signal    complement(5232..5353)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5455..5479
                     /label=ConE scar
                     /note="ConE scar"
     promoter        5499..5828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        5876..5923
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             5942..6538
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     CDS             6545..7537
                     /label=Delta-TK
                     /note="truncated herpes simplex virus thymidine kinase
                     (Salomon et al., 1995)"
     promoter        complement(7567..7585)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    7746..7867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   8064..8126
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac right (3') inverted repeat"
     regulatory      complement(8175..8201)
                     /label=KanR terminator
                     /note="KanR terminator"
                     /regulatory_class="terminator"
     CDS             complement(8205..9017)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(9018..9165)
                     /label=KanR promoter
                     /note="KanR promoter"
                     /regulatory_class="promoter"
     rep_origin      9242..9830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"