pDG2i-Flag-Puro vector (V008019)

LOCUS       V008019                 9846 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008019
VERSION     V008019
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9846)
  AUTHORS   De Groote P, Grootjans S, Lippens S, Eichperger C, Leurs K, Kahr I,
            Tanghe G, Bruggeman I, De Schamphelaire W, Urwyler C, Vandenabeele
            P, Haustraete J, Declercq W.
  TITLE     Generation of a new Gateway-compatible inducible lentiviral vector
            platform allowing easy derivation of co-transduced cells
  JOURNAL   BioTechniques 60 (5), 252-259 (2016)
   PUBMED   27177818
REFERENCE   2  (bases 1 to 9846)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-DEC-2015) BCCM/LMBP, Universiteit Gent,
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 9846)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9846)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "BioTechniques"; date: "2016"; volume: "60"; issue: "5"; pages:
            "252-259"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-DEC-2015) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
            9052, BELGIUM"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    75..90
                     /label="HIV-1 cPPT"
                     /note="HIV-1 cPPT"
     LTR             92..325
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    397..531
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      558..693
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(714..732)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(742..758)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      900..1355
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1381..1485
                     /label="AmpR promoter"
     CDS             1486..2343
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2517..3105
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3393..3414
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3429..3459
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3467..3483
                     /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)."
     primer_bind     3491..3507
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3528..3546
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3574..3800
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3801..3981
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4028..4153
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    4646..4879
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5064..5108
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             5257..5298
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     protein_bind    5396..5414
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5431..5449
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5467..5485
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5503..5521
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5538..5556
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5574..5592
                     /gene="tetO"
                     /label="tet operator"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5604..5642
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    5707..5831
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5856..5886
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             5940..6596
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             6941..7243
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(7287..7411)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7455..7478
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     promoter        7523..7853
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             7905..8609
                     /label="rtTA3"
                     /note="reverse tetracycline transactivator 3"
     protein_bind    complement(8613..8646)
                     /label="loxP511"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    8669..9251
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             9250..9843
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"