Basic Vector Information
- Vector Name:
- pLV3-8×GTIIC-EGFP-Fluc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8714 bp
- Type:
- Transcriptional activation
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- hPGK
- Growth Temperature:
- 37℃
pLV3-8×GTIIC-EGFP-Fluc vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pLV3-8×GTIIC-EGFP-Fluc vector Sequence
LOCUS 62056_14950 8714 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8714)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8714
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 7..507
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 522..2171
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
LTR 2245..2478
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2550..2684
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 2711..2846
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(2867..2885)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2895..2911)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3053..3508
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3534..3638
/label=AmpR promoter
CDS 3639..4496
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4670..5258
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5546..5567
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5582..5612
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5620..5636
/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 5644..5660
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 5681..5699
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 5727..5953
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 6181..6306
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6799..7032
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7217..7261
/codon_start=1
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
/translation="KNEQELLELDKWASL"
primer_bind 7619..7635
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 7787..8503
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 8545..8662
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 8711..8714
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
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