Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000612 | pSpliceExpress | 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:
- pSpliceExpress
- Antibiotic Resistance:
- Chloramphenicol, Ampicillin
- Length:
- 6920 bp
- Copy Number:
- High Copy
- Promoter:
- SV40
pSpliceExpress 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.
pSpliceExpress vector Sequence
LOCUS Exported 6920 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSpliceExpress
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6920)
AUTHORS Kishore S, Khanna A, Stamm S
TITLE Rapid generation of splicing reporters with pSpliceExpress.
JOURNAL Gene. 2008 Dec 31;427(1-2):104-10. Epub 2008 Oct 1.
PUBMED 18930792
REFERENCE 2 (bases 1 to 6920)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene.";
date: "2008-12-31"; volume: "427(1-2)"; pages: "104-10. Epub 2008
Oct 1"
FEATURES Location/Qualifiers
source 1..6920
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..112
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 161..183
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 175..192
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 176..192
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 209..440
/gene="mutant version of attP"
/label=attP1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
CDS complement(836..1141)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(938..957)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS complement(1491..2144)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
primer_bind 2059..2078
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2145..2247)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(2392..2623)
/gene="mutant version of attP"
/label=attP2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
primer_bind complement(2641..2660)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2642..2660)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2665..2681)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2665..2681)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3302..3320)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3388..3492
/gene="bla"
/label=AmpR promoter
CDS 3493..4353
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3711..3730)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
primer_bind 4889..4908
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter complement(5463..5659)
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5477..5612
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5531..5550)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 5767..5786
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
terminator 6827..6854
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"