Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017124 | TCRalpha_E2A_SP-SNAPf-TCRbeta_P2A_CD3e_P2A_CD3zeta | 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:
- TCRalpha_E2A_SP-SNAPf-TCRbeta_P2A_CD3e_P2A_CD3zeta
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12681 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Promoter:
- SFFV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
TCRalpha_E2A_SP-SNAPf-TCRbeta_P2A_CD3e_P2A_CD3zeta 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.
TCRalpha_E2A_SP-SNAPf-TCRbeta_P2A_CD3e_P2A_CD3zeta vector Sequence
LOCUS V017124 12681 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017124
VERSION V017124
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 12681)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..12681
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 17..605
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 851..1180
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 2314..2379
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2509..2529
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 2954..3088
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 3256..3889
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 3936..4061
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4558..4791
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 4976..5020
/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 5169..5210
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 5305..5422
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 5575..5982
/label="SFFV promoter"
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
CDS 6063..6401
/gene="TRAV8-4"
/label="T cell receptor alpha variable 8-4"
/note="T cell receptor alpha variable 8-4 from Homo
sapiens. Accession#: P01737"
CDS 6474..6893
/gene="TRAC"
/label="T cell receptor alpha chain constant"
/note="T cell receptor alpha chain constant from Homo
sapiens. Accession#: P01848"
CDS 6909..6968
/label="E2A"
/note="2A peptide from equine rhinitis A virus polyprotein"
CDS 7044..7586
/label="SNAP-tag(R)"
/note="human O6-alkylguanine-DNA-alkyltransferase variant
that forms covalent bonds with benzylguanine derivatives"
CDS 7944..8471
/gene="TRBC1"
/label="T cell receptor beta constant 1"
/note="T cell receptor beta constant 1 from Homo sapiens.
Accession#: P01850"
CDS 8487..8543
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 8550..9170
/gene="CD3E"
/label="T-cell surface glycoprotein CD3 epsilon chain"
/note="T-cell surface glycoprotein CD3 epsilon chain from
Homo sapiens. Accession#: P07766"
CDS 9186..9239
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 9246..9737
/gene="CD247"
/label="T-cell surface glycoprotein CD3 zeta chain"
/note="T-cell surface glycoprotein CD3 zeta chain from Homo
sapiens. Accession#: P20963"
CDS complement(10271..10282)
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
LTR 10498..10731
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(10756..10774)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
promoter 11562..11666
/label="AmpR promoter"
CDS 11667..12524
/label="AmpR"
/note="beta-lactamase"