Basic Vector Information
- Vector Name:
- pLX-TRE-dCas9-KRAB-MeCP2-BSD
- Antibiotic Resistance:
- Ampicillin
- Length:
- 16734 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Blast
- Promoter:
- TRE3G
- Growth Temperature:
- 30℃
pLX-TRE-dCas9-KRAB-MeCP2-BSD 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 ℃.
pLX-TRE-dCas9-KRAB-MeCP2-BSD vector Sequence
LOCUS 62056_16345 16734 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16734)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..16734
/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 2050..2167
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2929..3307
/label=TRE3G promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G"
CDS 3340..7443
/label=Cas9m4
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS 7579..7764
/label=KRAB
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
CDS 7798..7818
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 7828..8685
/label=MeCP2
/note="transcriptional repression domain from rat
methyl-CpG-binding protein 2 (Nan et al., 1997)"
polyA_signal 8784..8832
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
promoter 9063..9274
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 9320..10063
/label=Tet-On(R) 3G
/note="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
CDS 10064..10117
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 10139..10534
/label=BSD
/note="blasticidin S deaminase"
misc_feature 10598..11186
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 11296..11529
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(11554..11572)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 12360..12464
/label=AmpR promoter
CDS 12465..13322
/label=AmpR
/note="beta-lactamase"
rep_origin 13496..14084
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 14330..14659
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
intron 15793..15858
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 15988..16008
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 16433..16567
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
This page is informational only.