Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012075 | pBS-L1PA1-CH-mneo | 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:
- pBS-L1PA1-CH-mneo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11342 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Promoter:
- SV40
- 5' Primer:
- T7
- 3' Primer:
- SP6
pBS-L1PA1-CH-mneo 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.
pBS-L1PA1-CH-mneo vector Sequence
LOCUS V012075 11342 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012075
VERSION V012075
KEYWORDS pBS-L1PA1-CH-mneo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11342)
AUTHORS Wagstaff BJ, Barnerssoi M, Roy-Engel AM
TITLE Evolutionary conservation of the functional modularity of primate
and murine LINE-1 elements.
JOURNAL PLoS One. 2011 May 10;6(5):e19672. doi:
10.1371/journal.pone.0019672.
PUBMED 21572950
REFERENCE 2 (bases 1 to 11342)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11342)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
date: "2011-05-10"; pages: "
10.1371/journal.pone.0019672"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11342
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 4..459
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 585..607
/label="M13/pUC Forward"
/note="In lacZ gene"
primer_bind 599..616
/label="M13 Forward"
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 600..616
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
enhancer 935..1314
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 1315..1518
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1515..1539
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS 1580..2593
/gene="L1RE1"
/label="LINE-1 retrotransposable element ORF1 protein"
/note="LINE-1 retrotransposable element ORF1 protein from
Homo sapiens. Accession#: Q9UN81"
CDS 2660..6484
/note="LINE-1 retrotransposable element ORF2 protein from
Homo sapiens. Accession#: O00370"
/label="LINE-1 retrotransposable element ORF2 protein"
primer_bind complement(6655..6674)
/label="Neo-F"
/note="Neomycin resistance gene, forward primer"
primer_bind 8166..8185
/label="Neo-R"
/note="Neomycin resistance gene, reverse primer"
promoter complement(8295..8624)
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
polyA_signal complement(8729..8863)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(9154..9172)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9193..9209)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9193..9209)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9206..9228)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 9217..9233
/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(9241..9271)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(9286..9307)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9424..9441)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(9595..10183)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10357..11214)
/label="AmpR"
/note="beta-lactamase"
promoter complement(11215..11319)
/label="AmpR promoter"