pLXSN-HXY-TCR vector (V013994)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013994 pLXSN-HXY-TCR In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pLXSN-HXY-TCR
Antibiotic Resistance:
Ampicillin
Length:
7676 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Retrovirus
Selection Marker:
Neo/G418
Promoter:
SV40
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pLXSN-HXY-TCR vector Map

pLXSN-HXY-TCR7676 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075005' LTRMMLV Psigag (truncated)T cell receptor beta variable 11-2P2AT cell receptor alpha variable 17T-cell receptor alpha chain constantSV40 promoterNeoR/KanRLTRbomoriAmpRAmpR promoter

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.

pLXSN-HXY-TCR vector Sequence

LOCUS       V013994                 7676 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013994
VERSION     V013994
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7676)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2..589
                     /label="5' LTR"
                     /note="long terminal repeat from Moloney murine sarcoma
                     virus"
     misc_feature    652..851
                     /label="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1052..1468
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     CDS             1482..1826
                     /gene="TRBV11-2"
                     /label="T cell receptor beta variable 11-2"
                     /note="T cell receptor beta variable 11-2 from Homo
                     sapiens. Accession#: A0A584"
     CDS             2421..2477
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             2478..2813
                     /gene="TRAV17"
                     /label="T cell receptor alpha variable 17"
                     /note="T cell receptor alpha variable 17 from Homo sapiens.
                     Accession#: A0A0B4J275"
     CDS             2877..3284
                     /gene="Trac"
                     /label="T-cell receptor alpha chain constant"
                     /note="T-cell receptor alpha chain constant from Mus
                     musculus. Accession#: P01849"
     promoter        3305..3634
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3694..4485
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     LTR             4548..5141
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    5352..5492
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5678..6266)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6440..7297)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7298..7402)
                     /label="AmpR promoter"