p16L1L2 vector (V016696)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016696 p16L1L2 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:
p16L1L2
Antibiotic Resistance:
Bleomycin
Length:
6388 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Promoter:
EF-1α core
Growth Strain(s):
DH5a
Growth Temperature:
37℃

p16L1L2 vector Map

p16L1L26388 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300SV40 promoterSV40 intronMinor capsid protein L2SV40 poly(A) signalEF-1-alpha core promoter5' LTR (truncated)Major capsid protein L1EF-1-alpha poly(A) signaloriEM7 promoterBleoR

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.

p16L1L2 vector Sequence

LOCUS       V016696                 6388 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016696
VERSION     V016696
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6388)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..342
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     intron          415..511
                     /label="SV40 intron"
                     /note="modified SV40 intron with splice donor and acceptor
                     sites"
     CDS             581..1999
                     /gene="L2"
                     /label="Minor capsid protein L2"
                     /note="Minor capsid protein L2 from Human papillomavirus
                     type 16. Accession#: P03107"
     polyA_signal    complement(2065..2186)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        2387..2598
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2611..2879
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2911..4425
                     /gene="L1"
                     /label="Major capsid protein L1"
                     /note="Major capsid protein L1 from Human papillomavirus
                     type 16. Accession#: P03101"
     polyA_signal    4477..5049
                     /label="EF-1-alpha poly(A) signal"
                     /note="human EF-1-alpha polyadenylation signal"
     rep_origin      5220..5808
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5894..5941
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             5961..6332
                     /label="BleoR"
                     /note="antibiotic-binding protein"