Basic Vector Information
- Vector Name:
- pMOS013: mito-roGFP2-Grx1 glutathione redox sensor (mitochondrial)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9003 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Blast
- Promoter:
- hPGK
- Growth Temperature:
- 30℃
pMOS013: mito-roGFP2-Grx1 glutathione redox sensor (mitochondrial) 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 ℃.
pMOS013: mito-roGFP2-Grx1 glutathione redox sensor (mitochondrial) vector Sequence
LOCUS 62056_17215 9003 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 9003)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9003
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 228..353
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 846..1079
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1264..1308
/codon_start=1
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
/translation="KNEQELLELDKWASL"
promoter 1483..1983
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 1995..2390
/codon_start=1
/label=BSD
/note="blasticidin S deaminase"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 2450..2567
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2684..2987
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2988..3191
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 3214..3238
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 3341..4054
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
protein_bind complement(4520..4544)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
CDS 4546..4587
/codon_start=1
/label=V5 tag
/note="epitope tag from simian virus 5"
/translation="GKPIPNPLLGLDST"
misc_feature 4629..5217
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5289..5522
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5600..5734
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 5761..5896
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(5917..5935)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5945..5961)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6103..6558
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6584..6688
/label=AmpR promoter
CDS 6689..7546
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 7720..8308
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 8596..8617
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8632..8662
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8670..8686
/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 8694..8710
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 8731..8749
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 8777..9003
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
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