Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010447 | pLMPd-Ametrine | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pLMPd-Ametrine is a retroviral vector with an mAmetrine marker, for expressing a short hairpin RNA (shRNA) with an "UltramiR" microRNA scaffold.
- Vector Name:
- pLMPd-Ametrine
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6740 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- transOMIC
- Copy Number:
- High copy number
- Promoter:
- MSCV
- Growth Strain(s):
- Top10
- Growth Temperature:
- 37℃
pLMPd-Ametrine vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Li C, Mori LP, Lyu S, Bronson R, Getzler AJ, Pipkin ME, Valente ST. The chaperone protein p32 stabilizes HIV-1 Tat and strengthens the p-TEFb/RNAPII/TAR complex promoting HIV transcription elongation. Proc Natl Acad Sci U S A. 2023 Jan 3;120(1):e2217476120.
pLMPd-Ametrine vector Sequence
LOCUS pLMPd-Ametrine. 6740 bp DNA circular SYN 01-JAN-1980
DEFINITION Retroviral vector with an mAmetrine marker, for expressing a short
hairpin RNA (shRNA) with an """"UltramiR"""" microRNA scaffold.
ACCESSION .
VERSION .
KEYWORDS pLMPd-Ametrine
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6740)
AUTHORS transOMIC
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6740)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6740
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..515
/label=MSCV
/note="Murine embryonic stem cell virus promoter including
the enhancer and promoter region of PCMV virus and 5'
untranslated region of dl-587rev retrovirus"
misc_feature 579..920
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
ncRNA 1443..1564
/label=5' miR-30a
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
misc_RNA 1565..1633
/label=shRNA
/note="shRNA"
/note="short hairpin RNA"
gap 1568..1589
/estimated_length=22
gap 1609..1630
/estimated_length=22
ncRNA 1634..1760
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
promoter 1774..2273
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 2294..3010
/label=mAmetrine1.1
/note="photostable yellow fluorescent protein with a large
Stokes shift, excitable with violet light (Ai et al.,
2008)"
LTR 3102..3616
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
primer_bind complement(3785..3801)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3809..3825)
/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)."
promoter complement(3833..3863)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3878..3899)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4187..4775)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4949..5806)
/label=AmpR
/note="beta-lactamase"
promoter complement(5807..5911)
/label=AmpR promoter
LTR 6739..6740
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"