Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007516 | pJBEI-6409 | 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:
- pJBEI-6409
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 14932 bp
- Type:
- Bacterial Expression, Synthetic Biology ; BglBrick
- Replication origin:
- p15A ori
- Copy Number:
- High Copy
- Promoter:
- PlacUV5
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- F5
- 3' Primer:
- R1
pJBEI-6409 vector 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.
pJBEI-6409 vector Sequence
LOCUS V007516 14932 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007516
VERSION V007516
KEYWORDS pJBEI-6409
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 14932)
AUTHORS Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
CJ, Lee TS
TITLE Metabolic engineering of Escherichia coli for limonene and perillyl
alcohol production.
JOURNAL Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
PUBMED 23727191
REFERENCE 2 (bases 1 to 14932)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 14932)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Metab
Eng."; date: "2013-05-29"; pages: "
10.1016/j.ymben.2013.05.004"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14932
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(15..36)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(52..1131)
/label="lacI"
/note="lac repressor"
promoter complement(1132..1209)
/label="lacI promoter"
primer_bind 1415..1434
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 1520..1550
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 1558..1574
/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)."
CDS 1610..2791
/gene="atoB"
/label="Acetyl-CoA acetyltransferase"
/note="Acetyl-CoA acetyltransferase from Escherichia coli
(strain K12). Accession#: P76461"
terminator 5324..5395
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5411..5438
/label="T7Te terminator"
/note="phage T7 early transcription terminator"
promoter 5650..5679
/label="trc promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 5687..5703
/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)."
CDS 5729..7057
/gene="ERG12"
/label="Mevalonate kinase"
/note="Mevalonate kinase from Saccharomyces cerevisiae
(strain ATCC 204508 / S288c). Accession#: P07277"
CDS 7081..8433
/gene="ERG8"
/label="Phosphomevalonate kinase"
/note="Phosphomevalonate kinase from Saccharomyces
cerevisiae (strain ATCC 204508 / S288c). Accession#:
P24521"
CDS 8457..9644
/gene="MVD1"
/label="Diphosphomevalonate decarboxylase"
/note="Diphosphomevalonate decarboxylase from Saccharomyces
cerevisiae (strain ATCC 204508 / S288c). Accession#:
P32377"
CDS 9668..10213
/gene="idi"
/label="Isopentenyl-diphosphate Delta-isomerase"
/note="Isopentenyl-diphosphate Delta-isomerase from
Escherichia coli (strain K12 / DH10B). Accession#: B1XEH6"
promoter 10432..10461
/label="trc promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 10469..10485
/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)."
terminator 13097..13168
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 13184..13211
/label="T7Te terminator"
/note="phage T7 early transcription terminator"
primer_bind complement(13239..13256)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(13373..13918)
/direction=LEFT
/label="p15A ori"
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
terminator complement(14032..14126)
/label="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS complement(14150..14806)
/label="CmR"
/note="chloramphenicol acetyltransferase"
promoter complement(14807..14909)
/label="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"