pULTRA-CNF vector (V017396)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017396 pULTRA-CNF 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:
pULTRA-CNF
Antibiotic Resistance:
Streptomycin
Length:
4977 bp
Type:
Protein expression
Replication origin:
CloDF13 ori
Host:
E. coli
Promoter:
tac
Growth Temperature:
37℃

pULTRA-CNF vector Map

pULTRA-CNF4977 bp6001200180024003000360042004800tac promoterlac operatorTyrosine--tRNA ligaseSmRAmpR promoterCloDF13 oriCAP binding sitelacIlacI 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.

pULTRA-CNF vector Sequence

LOCUS       V017396                 4977 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017396
VERSION     V017396
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4977)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..39
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    47..63
                     /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)."
     CDS             87..1004
                     /gene="tyrS"
                     /label="Tyrosine--tRNA ligase"
                     /note="Tyrosine--tRNA ligase from Methanocaldococcus
                     jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM
                     10045 / NBRC 100440). Accession#: Q57834"
     CDS             complement(1904..2692)
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(2693..2784)
                     /label="AmpR promoter"
     rep_origin      complement(2832..3570)
                     /direction=LEFT
                     /label="CloDF13 ori"
                     /note="Plasmids containing the CloDF13 (CDF) origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(3746..3767)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3783..4862)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4863..4940)
                     /label="lacI promoter"