Basic Vector Information
3rd gen lentiviral empty control vector, Puro
- Vector Name:
- pLenti X1 Puro empty (w610-1)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7424 bp
- Type:
- Lentiviral Expression Plasmids
- Replication origin:
- ori
- Promoter:
- mPGK
pLenti X1 Puro empty (w610-1) 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 ℃.
pLenti X1 Puro empty (w610-1) vector Sequence
LOCUS 40924_27997 7424 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7424)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7424)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7424
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 27..131
/label=AmpR promoter
CDS 132..989
/label=AmpR
/note="beta-lactamase"
rep_origin 1163..1751
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2039..2060
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2075..2105
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2113..2129
/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 2137..2153
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2174..2192
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2220..2447
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 2448..2628
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2672..2797
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3290..3523
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3707..3751
/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 3900..3941
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 4012..4129
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 4161..4185
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(4251..4275)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 4306..4894
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
promoter 4908..5407
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 5428..6024
/label=PuroR
/note="puromycin N-acetyltransferase"
LTR 6165..6398
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6470..6604
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 6610..6745
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(6778..6796)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6806..6822)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6967..7422
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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