Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007336 | MSGV Hu Acceptor PGK-NGFR | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- MSGV Hu Acceptor PGK-NGFR
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8069 bp
- Type:
- Retroviral
- Replication origin:
- ori
- Selection Marker:
- tNGFR
- Copy Number:
- High Copy
- Promoter:
- MSCV
- Cloning Method:
- Restriction Enzyme
MSGV Hu Acceptor PGK-NGFR 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.
MSGV Hu Acceptor PGK-NGFR vector Sequence
LOCUS V007336 8069 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007336
VERSION V007336
KEYWORDS MSGV Hu Acceptor PGK-NGFR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8069)
AUTHORS Coren LV, Jain S, Trivett MT, Ohlen C, Ott DE
TITLE Production of retroviral constructs for effective transfer and
expression of T-cell receptor genes using Golden Gate cloning.
JOURNAL Biotechniques. 2015 Mar 1;58(3):135-9. doi: 10.2144/000114265.
eCollection 2015.
PUBMED 25757546
REFERENCE 2 (bases 1 to 8069)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8069)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.2144/000114265";
journalName: "Biotechniques"; date: "2015-03-1- 1"; volume: "58";
issue: "3"; pages: "135-9"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8069
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 343..518
/label="5' LTR (truncated)"
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
CDS 1021..1428
/label="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
misc_feature 1438..1810
/label="pol region"
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
CDS 1852..2271
/gene="TRAC"
/label="T cell receptor alpha chain constant"
/note="T cell receptor alpha chain constant from Homo
sapiens. Accession#: P01848"
CDS 2368..2895
/gene="TRBC1"
/label="T cell receptor beta constant 1"
/note="T cell receptor beta constant 1 from Homo sapiens.
Accession#: P01850"
promoter 2911..3409
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
misc_feature 4290..4342
/label="MCS"
/note="multiple cloning site"
LTR 4386..4900
/label="3' LTR"
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind complement(5072..5088)
/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(5096..5126)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(5141..5162)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5279..5296)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(5450..6038)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6212..7069)
/label="AmpR"
/note="beta-lactamase"
promoter complement(7070..7174)
/label="AmpR promoter"
primer_bind 7242..7260
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7298..7320)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 7420..7439
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7648..7664
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7863..7882
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"