Basic Vector Information
- Vector Name:
- pDG2i-Flag-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9846 bp
- Type:
- Lentiviral vector
- Replication origin:
- ori
- Source/Author:
- 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.
- Promoter:
- RSV
pDG2i-Flag-Puro vector Vector Map
pDG2i-Flag-Puro vector Sequence
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"
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