pKF-P14MMP2AG-T2APuroR-5h vector (V016240)

LOCUS       62056_13380        6855 bp DNA     circular SYN 07-JUL-2019
DEFINITION  Cloning vector pKF-P14MMP2AG-T2APuroR-5h, complete sequence.
ACCESSION   MK816962
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6855)
  AUTHORS   Farquhar KS, Charlebois DA, Szenk M, Cohen J, Nevozhay D, Balazsi G.
  TITLE     Role of network-mediated stochasticity in mammalian drug resistance
  JOURNAL   Nat Commun 10 (1), 2766 (2019)
  PUBMED    31235692
REFERENCE   2  (bases 1 to 6855)
  AUTHORS   Farquhar KS, Charlebois DA, Szenk M, Cohen J, Nevozhay D, Balazsi G.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2019) Laufer Center for Physical and Quantitative 
            Biology, Stony Brook University, 100 Nicolls Road, Stony Brook, NY 
            11790, USA
REFERENCE   3  (bases 1 to 6855)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2019"; volume: "10"; issue: "1"; pages: "2766"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2019) Laufer Center for Physical and Quantitative Biology, 
            Stony Brook University, 100 Nicolls Road, Stony Brook, NY 11790, 
            USA"
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        257..571
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     CDS             576..1319
                     /label=rtTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
     misc_feature    1329..1385
                     /note="eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides; P2A; from porcine teschovirus 1 polyprotein"
     CDS             1386..2102
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    2103..2156
                     /note="eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides; T2A; from Thosea asigna virus capsid protein"
     misc_feature    2157..2756
                     /note="confers resistance to puromycin; PuroR; puromycin 
                     N-acetyltransferase"
     polyA_signal    2813..3037
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3321..3368
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3376..4395
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    4528..4661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4698..4714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4722..4738)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4746..4776)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4791..4812)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5862..6719)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6720..6824)
                     /label=AmpR promoter