Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005293 | pJN105 | 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:
- pJN105
- Antibiotic Resistance:
- Gentamicin
- Length:
- 6055 bp
- Type:
- Expression vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Newman JR, Fuqua C.
- Promoter:
- araBAD
pJN105 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.
pJN105 vector Sequence
LOCUS Exported 6055 bp DNA circular SYN 05-AUG-2024
DEFINITION Expression vector pJN105, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6055)
AUTHORS Newman JR, Fuqua C.
TITLE Broad-host-range expression vectors that carry the
L-arabinose-inducible Escherichia coli araBAD promoter and the araC
regulator
JOURNAL Gene 227 (2), 197-203 (1999)
PUBMED 10023058
REFERENCE 2 (bases 1 to 6055)
AUTHORS Newman JR, Fuqua C.
TITLE Direct Submission
JOURNAL Submitted (21-AUG-1999) Biology, Indiana University, JH142, 1001 E.
3rd St., Bloomington, IN 47405-6801, USA
REFERENCE 3 (bases 1 to 6055)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6055)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6055)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1999"; volume: "227"; issue: "2"; pages: "197-203"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-AUG-1999) Biology, Indiana University, JH142, 1001 E. 3rd St.,
Bloomington, IN 47405-6801, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6055
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(5810..6055,1..5809)
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(18..548)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(737..765)
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 1035..1056
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1071..1101
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1109..1125
/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)."
primer_bind 1133..1149
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 1170..1188
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 1218..1234
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(1329..2204)
/codon_start=1
/label=araC
/note="L-arabinose regulatory protein"
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 2231..2515
/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)"
primer_bind complement(2555..2571)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2604..2622)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2632..2648)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3358..4017)
/codon_start=1
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
rep_origin complement(4018..4787)
/direction=LEFT
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
promoter 4892..4943
/label=promoter for mob
misc_feature 4910..4932
/label=RSA
/note="transfer origins (also called recombination site A
[RSA]), is known as the specific site necessary for
mobilization and recombination mediated by a Mob/Pre
protein. "
CDS 5011..6015
/codon_start=1
/product="mob encodes the plasmid mobilization functions
that allow conjugal delivery of the plasmids into a variety
of bacteria from E. coli strains harboring the RK2 conjugal
transfer functions."
/label=mob
/translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLAAL"