Basic Vector Information
- Vector Name:
- CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13286 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Selection Marker:
- zsGreen1
- Promoter:
- EF-1α
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- NA
- 3' Primer:
- NA
CAPTURE2_pLVX-EF1a-dCas9-CBio-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 ℃.
CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1 vector Sequence
LOCUS CAPTURE2_pLVX-EF 13286 bp DNA circular SYN 13-MAY-2021
DEFINITION CAPTURE2.0 vector containing dCas9-CBio, IRES and zsGreen1.
ACCESSION .
VERSION .
KEYWORDS CAPTURE2_pLVX-EF1a-dCas9-CBio-IRES-zsGreen1.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13286)
AUTHORS Liu X, Chen Y, Zhang Y, Liu Y, Liu N, Botten GA, Cao H, Orkin SH,
Zhang MQ, Xu J
TITLE Multiplexed capture of spatial configuration and temporal dynamics
of locus-specific 3D chromatin by biotinylated dCas9.
JOURNAL Genome Biol. 2020 Mar 5;21(1):59. doi: 10.1186/s13059-020-01973-w.
PUBMED 32138752
REFERENCE 2 (bases 1 to 13286)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13286)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genome
Biol."; date: "2020-03-5"; pages: "
10.1186/s13059-020-01973-w"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13286
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2027..2144
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2338..3519
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
regulatory 3558..3567
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3573..3593
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3591..7694
/label=dCas9
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS 7704..7724
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
misc_feature 7972..8551
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 8546..9238
/label=ZsGreen1
/note="Zoanthus green fluorescent protein"
misc_feature 9255..9843
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 10050..10683
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(10811..10827)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10811..10827)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10824..10846)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10835..10851
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(10859..10889)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(10904..10925)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11042..11059)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11213..11801)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11975..12832)
/label=AmpR
/note="beta-lactamase"
promoter complement(12833..12937)
/label=AmpR promoter
polyA_signal 12985..13119
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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