pTarget-eGFP-iFlp-neo vector (V015562)

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LOCUS       62056_20975        7482 bp DNA     circular SYN 24-MAY-2021
DEFINITION  Cloning vector pTarget-eGFP-iFlp-neo, complete sequence.
ACCESSION   MW246026
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7482)
  AUTHORS   Dias MM, Vidigal J, Sequeira DP, Alves PM, Teixeira AP, Roldao A.
  TITLE     Insect High FiveTM cell line development using site-specific flipase
            recombination technology
  JOURNAL   G3 (Bethesda) (2021) In press
  PUBMED    33982066
REFERENCE   2  (bases 1 to 7482)
  AUTHORS   Dias MM, Vidigal J, Sequeira DP, Alves PM, Teixeira AP, Roldao A.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2020) Animal Cell Technology Unit, iBET, Instituto
            de Biologia Experimental e Tecnologica, Av. da Republica, Qta. do 
            Marques, ITQB/iBET building, Oeiras 2780-157, Portugal
REFERENCE   3  (bases 1 to 7482)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda) (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-2020) Animal Cell Technology Unit, iBET, Instituto de 
            Biologia Experimental e Tecnologica, Av. da Republica, Qta. do 
            Marques, ITQB/iBET building, Oeiras 2780-157, Portugal"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..48
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        207..225
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             253..969
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             1075..1116
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             1126..1143
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    1161..1290
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     primer_bind     1432..1448
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        1469..1951
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        1955..2546
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus
                     Autographa californica"
     sig_peptide     2552..2611
                     /label=IgM signal sequence
                     /note="signal sequence from mouse IgM heavy chain"
     CDS             2624..2647
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             2654..2668
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             2686..3981
                     /label=FLPo
                     /note="nuclear-targeted site-specific recombinase"
     CDS             4037..4063
                     /label=9xHis
                     /note="9xHis affinity tag"
     terminator      4085..4392
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     regulatory      complement(4505..4553)
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(4620..5411)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_recomb     5419..5466
     promoter        5515..5619
                     /label=AmpR promoter
     misc_feature    5804..6183
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     rep_origin      6652..7240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"