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pJW168 vector (V012456)

Price Information

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V012456 pJW168 In stock, instant shipping

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

Basic Vector Information

pSC101 ori is a low-copy replication origin that requires the Rep101 protein, which is a temperature-sensitive protein. When bacteria containing a plasmid with this ori is cultured at 37℃, the plasmid will be lost.

Vector Name:
pJW168
Antibiotic Resistance:
Ampicillin
Length:
5728 bp
Type:
Cloning Vectors
Replication origin:
pSC101 ori
Source/Author:
Lucigen
Copy Number:
Low copy number
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pJW168 vector Map

Copy Sequence View Map
pJW1685728 bp60012001800240030003600420048005400Crelac operatorlac UV5 promoterlac operatorlac UV5 promoterlacIq promoterlacICAP binding siteAmpR promoterAmpRpSC101 oriRep101(Ts)

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.

pJW168 vector Sequence

LOCUS       pJW168.        5728 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector allowing inducible expression of Cre recombinase 
            for in vivo site-specific recombination.
ACCESSION   .
VERSION     .
KEYWORDS    pJW168
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5728)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5728)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The temperature-sensitive pSC101 replicon enables pJW168 to be
FEATURES             Location/Qualifiers
     source          1..5728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..1035)
                     /label=Cre
                     /note="site-specific recombinase"
     protein_bind    1084..1100
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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(1108..1138)
                     /note="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1179..1195)
                     /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(1203..1233)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     promoter        1441..1518
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1519..2598
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2614..2635
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2755..2859
                     /label=AmpR promoter
     CDS             2860..3717
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4342..4564
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4612..5559
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"