Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017496 | pGERM | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pGERM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4807 bp
- Type:
- Bacteroides Plasmid
- Source/Author:
- Shoemaker,N.B., Wang,G.R. and Salyers,A.A.
- Copy Number:
- High copy number
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pGERM 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.
pGERM vector Sequence
LOCUS pGERM 4807 bp DNA circular SYN 26-JUL-2024
DEFINITION PGERM gene disruption vector, complete sequence.
ACCESSION EF155418
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4807)
AUTHORS Shoemaker NB, Wang GR, Salyers AA.
TITLE Multiple gene products and sequences required for excision of the
mobilizable integrated Bacteroides element NBU1
JOURNAL J. Bacteriol. 182 (4), 928-936 (2000)
PUBMED 10648516
REFERENCE 2 (bases 1 to 4807)
AUTHORS Chiang HC, Manchester JK, Gordon JI.
TITLE Regulation of nitrogen metabolism by a novel hybrid two-component
system protein in a prominent human gut symbiont, Bacteroides
thetaiotaomicron
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 4807)
AUTHORS Chiang HC, Manchester JK, Gordon JI.
TITLE Direct Submission
JOURNAL Submitted (03-DEC-2006) Center for Genome Sciences, Washington
University School of Medicine, 4444 Forest Park Ave. Campus Box
8510, Saint Louis, MO 63108, USA
REFERENCE 4 (bases 1 to 4807)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J.
Bacteriol."; date: "2000"; volume: "182"; issue: "4"; pages:
"928-936"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(03-DEC-2006) Center for Genome Sciences, Washington University
School of Medicine, 4444 Forest Park Ave. Campus Box 8510, Saint
Louis, MO 63108, USA"
FEATURES Location/Qualifiers
source 1..4807
/mol_type="other DNA"
/label=pGERM is pUC19 with the RK2 oriT and ermG
/note="pGERM is pUC19 with the RK2 oriT and ermG"
/db_xref="taxon:433620"
/organism="pGERM gene disruption vector"
CDS complement(44..367)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
DAA"
primer_bind 277..293
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 294..350
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(363..379)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 387..403
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(411..441)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 456..477
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(709..1080)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 1113..1224
/label=origin of transfer from 56 kb circular plasmid RK2
/note="origin of transfer from 56 kb circular plasmid RK2"
oriT 1113..1222
/note="incP origin of transfer"
rep_origin complement(1543..2131)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
gene complement(2302..3162)
/gene="AmpR"
/label=AmpR
CDS complement(2302..3162)
/codon_start=1
/transl_table=11
/gene="AmpR"
/product="beta lactamase"
/label=AmpR
/protein_id="ABO69572.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene 3650..4384
/gene="ermG"
/label=ermG
CDS 3650..4384
/codon_start=1
/transl_table=11
/gene="ermG"
/product="rRNA methyltransferase"
/label=ermG
/protein_id="ABO69573.1"
/translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
DINNISFEQFVSLFNSYKIFNG"