Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007552 | jk100 pSinEGdsp(p1:MAPPnL;p2:gfp:barcodelandingpad:4xboxB) | 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:
- jk100 pSinEGdsp(p1:MAPPnL;p2:gfp:barcodelandingpad:4xboxB)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13220 bp
- Type:
- Mammalian Expression, Synthetic Biology ; SIndbis
- Replication origin:
- ori
jk100 pSinEGdsp(p1:MAPPnL;p2:gfp:barcodelandingpad:4xboxB) 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.
jk100 pSinEGdsp(p1:MAPPnL;p2:gfp:barcodelandingpad:4xboxB) vector Sequence
LOCUS 40924_1434 13220 bp DNA circular SYN 13-MAY-2021
DEFINITION genomic Sindbis plasmid, encoding MAPPnL and a GFP RNA with a
barcode landing pad.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13220)
TITLE Zador Lab MAPseq plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 13220)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13220)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13220
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 7753..7800
/codon_start=1
/label=GFP11
/note="11th beta-strand of superfolder GFP"
/translation="RDHMVLHEYVNAAGIT"
CDS 7837..7866
/codon_start=1
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
CDS 7873..8418
/codon_start=1
/label=CLIP-tag(TM)
/note="human O6-alkylguanine-DNA-alkyltransferase variant
that forms covalent bonds with benzylcytosine derivatives"
/translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
VKEWLLAHEGHRLGKPGLG"
CDS 9193..9258
/codon_start=1
/label=lambda N peptide
/note="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/translation="MDAQTRRRERRAEKQAQWKAAN"
regulatory 9316..9325
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 9322..9387
/codon_start=1
/product="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/label=lambda N peptide
/translation="MDAQTRRRERRAEKQAQWKAAN"
regulatory 9445..9454
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 9451..9516
/codon_start=1
/product="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/label=lambda N peptide
/translation="MDAQTRRRERRAEKQAQWKAAN"
regulatory 9574..9583
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 9580..9645
/codon_start=1
/product="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/label=lambda N peptide
/translation="MDAQTRRRERRAEKQAQWKAAN"
CDS 9982..10698
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
promoter 11391..11495
/label=AmpR promoter
CDS 11496..12353
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 12527..13115
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"