Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007010 | pGRG37 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pGRG37
- Antibiotic Resistance:
- Chloramphenicol, Ampicillin
- Length:
- 14263 bp
- Type:
- Bacterial Expression
- Replication origin:
- pSC101 ori
- Copy Number:
- Low Copy
- Promoter:
- araBAD
- Cloning Method:
- Gateway Cloning
- 5' Primer:
- na
pGRG37 vector Map
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.
pGRG37 vector Sequence
LOCUS V007010 14263 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007010
VERSION V007010
KEYWORDS pGRG37
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 14263)
AUTHORS McKenzie GJ, Craig NL
TITLE Fast, easy and efficient: site-specific insertion of transgenes into
enterobacterial chromosomes using Tn7 without need for selection of
the insertion event.
JOURNAL BMC Microbiol. 2006 . 6():39.
PUBMED 16646962
REFERENCE 2 (bases 1 to 14263)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 14263)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC
Microbiol. 2006 . 6():39."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14263
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 6..129
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 155..185
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
CDS 239..916
/label="CmR"
/note="chloramphenicol acetyltransferase"
CDS 1239..1541
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(1585..1709)
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(2119..2994)
/label="araC"
/note="L-arabinose regulatory protein"
promoter 3021..3305
/label="araBAD promoter"
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 3423..4241
/gene="tnsA"
/label="Transposon Tn7 transposition protein TnsA"
/note="Transposon Tn7 transposition protein TnsA from
Escherichia coli. Accession#: P13988"
CDS 4231..6336
/gene="tnsB"
/label="Transposon Tn7 transposition protein TnsB"
/note="Transposon Tn7 transposition protein TnsB from
Escherichia coli. Accession#: P13989"
CDS 6336..8000
/gene="tnsC"
/label="Transposon Tn7 transposition protein TnsC"
/note="Transposon Tn7 transposition protein TnsC from
Escherichia coli. Accession#: P05846"
primer_bind complement(9996..10013)
/label="pBAD Reverse"
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 10166..10252
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 10344..10371
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 10390..10481
/label="AmpR promoter"
CDS 10482..11339
/label="AmpR"
/note="beta-lactamase"
primer_bind complement(11471..11490)
/label="F1ori-R"
/note="F1 origin, reverse primer"
oriT complement(11513..11622)
/direction=LEFT
/label="oriT"
/note="incP origin of transfer"
primer_bind complement(11660..11679)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
rep_origin 12362..12584
/label="pSC101 ori"
/note="low-copy replication origin that requires the Rep101
protein"
CDS 12632..13579
/label="Rep101(Ts)"
/note="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
mobile_element 14015..14180
/label="Tn7L"
/note="mini-Tn7 element (left end of the Tn7 transposon)"