Basic Vector Information
- Vector Name:
- Lenti_sgRNA(MS2)_MCP-KRAB-IRES-zsGreen1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12101 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- EF-1α
- Growth Temperature:
- 37℃
Lenti_sgRNA(MS2)_MCP-KRAB-IRES-zsGreen1 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)_MCP-KRAB-IRES-zsGreen1 vector Sequence
LOCUS V014960 12101 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014960
VERSION V014960
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12101)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12101
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..1179
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 1204..1224
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 1621..1815
/label="KRAB"
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
CDS 1882..1902
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
misc_feature 1944..2523
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 2518..3210
/label="ZsGreen1"
/note="Zoanthus green fluorescent protein"
protein_bind complement(3594..3627)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 3690..4278
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4803..4983
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 5015..5239
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 5285..5713
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5727..6056
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 6104..6151
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 6170..6541
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 6674..6807
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(6892..6922)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(6937..6958)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7246..7834)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8008..8865)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8866..8970)
/label="AmpR promoter"
enhancer 9236..9615
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 9616..9818
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 10002..11179
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 11289..11330
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 11438..11555
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 11605..11845
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 11891..11909
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
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