Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001167 | pSPL3 | 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
The pSPL3 vector is designed for exon trapping and splicing analysis. One of its key features is the presence of splice acceptor and donor sites that facilitate the detection of alternative splicing events. pSPL3 offers several advantages, such as high efficiency in capturing exons and providing insights into gene structure and function. It is ideal for researchers studying splicing regulation, identifying novel exons, and characterizing gene variants. When investigating complex splicing patterns or looking for potential disease-causing mutations related to splicing, pSPL3 is a valuable choice.
- Vector Name:
- pSPL3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6031 bp
- Type:
- Cloning vectors
- Replication origin:
- ori
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pSPL3 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.
References
- Pousada G, Lupo V, Cástro-Sánchez S, Álvarez-Satta M, Sánchez-Monteagudo A, Baloira A, Espinós C, Valverde D. Molecular and functional characterization of the BMPR2 gene in Pulmonary Arterial Hypertension. Sci Rep. 2017 May 15;7(1):1923. doi: 10.1038/s41598-017-02074-8. PMID: 28507310; PMCID: PMC5432510.
- Church DM, Stotler CJ, Rutter JL, Murrell JR, Trofatter JA, Buckler AJ. Isolation of genes from complex sources of mammalian genomic DNA using exon amplification. Nat Genet. 1994 Jan;6(1):98-105. doi: 10.1038/ng0194-98. PMID: 8136842.
pSPL3 vector Sequence
LOCUS Exported 6031 bp DNA circular SYN 21-SEP-2024
DEFINITION Exported.
ACCESSION V001167
VERSION .
KEYWORDS pSPL3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6031)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6031)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6031)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6031)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6031
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 10..339
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
misc_feature 693..698
/label=half BstXI
misc_feature 699..707
/label=SD
/note="donr site
"
CDS 715..957
/gene="vpu"
/label=Protein Vpu
/note="Protein Vpu from Human immunodeficiency virus type 1
group M subtype B (isolate BH10). Accession#: P69699"
CDS 875..3262
/codon_start=1
/product="gp160"
/label=HIV envelope protein gp160
/note="HIV gp160"
/translation="MRVKEKYQHLWRWGWRWGTMLLGMLMICSATEKLWVTVYYGVRDH
QNSGARAAAAGSQISGDPVPVWKEATTTLFCASDAKAYDTEVHNVWATHAGVPTDPNPQ
EVVLVNVTENFNMWKNDMVEQMHEDIISLWDQSLKPCVKLTPLCVSLKCTDLKNDTNTN
SSSGRMIMEKGEIKNCSFNISTSIRGKVQKEYAFFYKLDIIPIDNDTTSYTLTSCNTSV
ITQACPKVSFEPIPIHYCAPAGFAILKCNNKTFNGTGPCTNVSTVQCTHGIRPVVSTQL
LLNGSLAEEEVVIRSVNFTDNAKTIIVQLNTSVEINCTRPNNNTRKKIRIQRGPGRAFV
TIGKIGNMRQAHCNISRAKWNATLKQIASKLREQFGNNKTIIFKQSSGGDPEIVTHSFN
CGGEFFYCNSTQLFNSTWFNSTWSTEGSNNTEGSDTITLPCRIKQFINMWQEVGKAMYA
PPISGQIRCSSNITGLLLTRDGGNNNNGSEIFRPGGGDMRDNWRSELYKYKVVKIEPLG
VAPTKAKRRVVQREKRAVGIGALFLGFLGAAGSTMGAASMTLTVQARQLLSGIVQQQNN
LLRAIEAQQHLLQLTVWGIKQLQARILAVERYLKDQQLLGIWGCSGKLLCTTAVPWNAS
WSNKSLEQIWNHTTWMEWDREINNYTSLIHSLIEESQNQQEKNEQELLELDKWASLWNW
FNITNWLWYIKLFIMIVGGLVGLRIVFAVLSVVNRVRQGYSPLSFQTWRSPEGTRQARR
NRRRRWRERQRQIHFDQFTPQVQAAYQKVVAGVANALAHKYH"
misc_feature 3083..3094
/label=SA
/note="acceptor site"
misc_feature 3095..3100
/label=half BstXI
polyA_signal complement(3263..3397)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4071..4175
/label=AmpR promoter
CDS 4176..5033
/label=AmpR
/note="beta-lactamase"
rep_origin 5207..5795
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"