Basic Vector Information
- Vector Name:
- pLKO.1 GFP shRNA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7091 bp
- Type:
- Mammalian Expression, Lentiviral, RNAi
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- hPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- LKO.1 5'
pLKO.1 GFP shRNA 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 ℃.
pLKO.1 GFP shRNA vector Sequence
LOCUS pLKO.1_GFP_shRNA 7091 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pLKO.1 GFP shRNA.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7091)
AUTHORS Sancak Y, Peterson TR, Shaul YD, Lindquist RA, Thoreen CC, Bar-Peled
L, Sabatini DM
TITLE The Rag GTPases bind raptor and mediate amino acid signaling to
mTORC1.
JOURNAL Science. 2008 Jun 13. 320(5882):1496-501.
PUBMED 18497260
REFERENCE 2 (bases 1 to 7091)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7091)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.
2008 Jun 13. 320(5882):1496-501."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7091
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 23..42
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 276..293
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 410..431
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 446..476
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 484..500
/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 508..524
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 545..563
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 591..817
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 818..998
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1045..1170
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1663..1896
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2081..2125
/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 2274..2315
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
promoter 2423..2663
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 2775..2892
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2941..3451
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 3473..4069
/label=PuroR
/note="puromycin N-acetyltransferase"
LTR 4200..4433
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4505..4639
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 4666..4801
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(4822..4840)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4850..4866)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5008..5463
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5489..5593
/label=AmpR promoter
CDS 5594..6451
/label=AmpR
/note="beta-lactamase"
rep_origin 6625..7091
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
This page is informational only.