Basic Vector Information
- Vector Name:
- Lenti sgRNA(MS2)_Puro backbone
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10222 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Lenti sgRNA(MS2)_Puro backbone 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 ℃.
Lenti sgRNA(MS2)_Puro backbone vector Sequence
LOCUS V016656 10222 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V016656
VERSION V016656
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10222)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10222
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 86..415
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 463..510
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 529..900
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 1033..1166
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(1251..1281)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(1296..1317)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1605..2193)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2367..3224)
/label="AmpR"
/note="beta-lactamase"
promoter complement(3225..3329)
/label="AmpR promoter"
enhancer 3595..3974
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3975..4177
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 4419..4544
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5037..5270
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5455..5499
/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 5648..5689
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 5797..5914
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 5964..6204
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 6250..6268
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
promoter 6462..7640
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 7647..8243
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 8271..8859
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 9384..9564
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 9596..9820
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
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