Basic Vector Information
- Vector Name:
- pLenti SpBsmBI sgRNA Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8475 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Promoter:
- EF-1α
pLenti SpBsmBI sgRNA Puro 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 SpBsmBI sgRNA Puro vector Sequence
LOCUS pLenti_SpBsmBI_s 8475 bp DNA circular SYN 13-MAY-2021
DEFINITION An empty sgRNA expression vector for expression of sgRNA for Sp Cas9
(3rd generation lentiviral vector).
ACCESSION .
VERSION .
KEYWORDS pLenti SpBsmBI sgRNA Puro.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8475)
AUTHORS Pham H, Kearns NA, Maehr R
TITLE Transcriptional Regulation with CRISPR/Cas9 Effectors in Mammalian
Cells.
JOURNAL Methods Mol Biol. 2016;1358:43-57. doi: 10.1007/978-1-4939-3067-8_3.
PUBMED 26463376
REFERENCE 2 (bases 1 to 8475)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8475)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
Biol."; date: "2016"; pages: "
10.1007/978-1-4939-3067-8_3"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8475
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(20..38)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(59..75)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(83..99)
/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)."
promoter complement(107..137)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(152..173)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(290..307)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(461..1049)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1223..2080)
/label=AmpR
/note="beta-lactamase"
promoter complement(2081..2185)
/label=AmpR promoter
rep_origin complement(2211..2666)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2808..2824
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2834..2852
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(2873..3008)
/direction=LEFT
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal complement(3035..3169)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
LTR complement(3241..3474)
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature complement(3561..4149)
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4159..4755)
/label=PuroR
/note="puromycin N-acetyltransferase"
promoter complement(4778..5959)
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature complement(6164..6280)
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6311..6551
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 6579..6654
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS complement(6743..6784)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
CDS complement(6933..6977)
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(7162..7395)
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature complement(7888..8013)
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 8060..8240
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(8241..8467)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
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