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pQLinkG2 vector (V000735)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000735 pQLinkG2 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:
pQLinkG2
Antibiotic Resistance:
Ampicillin
Length:
5537 bp
Type:
Prokaryotic Expression Vectors
Replication origin:
ori
Promoter:
T5/lac

pQLinkG2 vector Map

Copy Sequence View Map
pQLinkG25537 bp60012001800240030003600420048005400T5 promoterlac operatorRBSGSTattB1TEV siteattB2lambda t0 terminatorCmRrrnB T1 terminatorCAP binding sitelacIlacIq promoterbomoriAmpRAmpR 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.

pQLinkG2 vector Sequence

LOCUS       40924_36213        5537 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5537)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5537)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..80
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    88..104
                     /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)."
     RBS             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             141..794
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     protein_bind    817..837
                     /label=attB1
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     CDS             840..860
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    complement(898..922)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      944..1038
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1132..1788
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      1856..1942
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(2001..2022)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2038..3117)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3118..3195)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    3387..3527
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3713..4301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4475..5332)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5333..5437)
                     /label=AmpR promoter