Basic Vector Information
- Vector Name:
- pLYS1-FLAG-MitoGFP-HA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8341 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- Low Copy
- Promoter:
- hPGK
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- custom
pLYS1-FLAG-MitoGFP-HA 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 ℃.
pLYS1-FLAG-MitoGFP-HA vector Sequence
LOCUS pLYS1-FLAG-MitoG 8341 bp DNA circular SYN 13-MAY-2021
DEFINITION Expression of GFP targeted to mitochondria with Flag and HA tags.
ACCESSION .
VERSION .
KEYWORDS pLYS1-FLAG-MitoGFP-HA.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8341)
AUTHORS Sancak Y, Markhard AL, Kitami T, Kovacs-Bogdan E, Kamer KJ, Udeshi
ND, Carr SA, Chaudhuri D, Clapham DE, Li AA, Calvo SE, Goldberger O,
Mootha VK
TITLE EMRE is an Essential Component of the Mitochondrial Calcium
Uniporter Complex.
JOURNAL Science. 2013 Nov 14.
PUBMED 24231807
REFERENCE 2 (bases 1 to 8341)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8341)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.
2013 Nov 14."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8341
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..227
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 228..408
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 455..580
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1073..1306
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1491..1535
/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 1684..1725
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
primer_bind 1833..1853
/label=hU6-F
/note="Human U6 promoter, forward primer"
promoter 2142..2345
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2560..2583
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 2614..3330
/label=EGFP
/note="enhanced GFP"
CDS 3358..3384
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
misc_feature 3435..3552
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 3601..4111
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 4133..4729
/label=PuroR
/note="puromycin N-acetyltransferase"
LTR 4860..5093
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5165..5299
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 5326..5461
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(5482..5500)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5510..5526)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5668..6123
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6149..6253
/label=AmpR promoter
CDS 6254..7111
/label=AmpR
/note="beta-lactamase"
rep_origin 7285..7873
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8027..8044
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 8161..8182
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8197..8227
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8235..8251
/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)."
primer_bind 8259..8275
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 8296..8314
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
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