Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008802 | pCAGGS-T7 | 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
- Vector Name:
- pCAGGS-T7
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7421 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Paterson RG, Russell CJ, Lamb RA.
- Promoter:
- CAG
pCAGGS-T7 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
- Jiang Y, Liu H, Liu P, Kong X. Plasmids driven minigenome rescue system for Newcastle disease virus V4 strain. Mol Biol Rep. 2009 Sep;36(7):1909-14. doi: 10.1007/s11033-008-9398-x. Epub 2008 Nov 15. PMID: 19011992.
pCAGGS-T7 vector Sequence
LOCUS Exported 7421 bp DNA circular SYN 02-SEP-2024
DEFINITION Exported.
ACCESSION V008802
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7421)
AUTHORS Paterson RG, Russell CJ, Lamb RA.
TITLE Fusion protein of the paramyxovirus SV5: destabilizing and
stabilizing mutants of fusion activation
JOURNAL Virology 270 (1), 17-30 (2000)
PUBMED 10772976
REFERENCE 2 (bases 1 to 7421)
AUTHORS Wickersham IR, Sullivan HA, Seung HS.
TITLE Production of glycoprotein-deleted rabies viruses for monosynaptic
tracing and high-level gene expression in neurons
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 7421)
AUTHORS Wickersham IR.
TITLE Direct Submission
JOURNAL Submitted (06-DEC-2009) Brain and Cognitive Sciences, Howard Hughes
Medical Institute and Massachusetts Institute of Technology, 43
Vassar St., Cambridge, MA 02139, USA
REFERENCE 4 (bases 1 to 7421)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 7421)
TITLE Direct Submission
REFERENCE 6 (bases 1 to 7421)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Virology";
date: "2000"; volume: "270"; issue: "1"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(06-DEC-2009) Brain and Cognitive Sciences, Howard Hughes Medical
Institute and Massachusetts Institute of Technology, 43 Vassar St.,
Cambridge, MA 02139, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT SGRef: number: 5; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7421
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 63..366
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 368..642
/label=chicken beta-actin promoter
intron 644..1659
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
CDS 1750..4398
/codon_start=1
/label=T7 RNA polymerase
/note="T7 RNA polymerase from Escherichia phage T7.
Accession#: P00573"
/translation="MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYE
MGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQ
FLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFK
KNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIE
STGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGI
TGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANV
ITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLE
FMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYY
WLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFE
YAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAK
KVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVM
TLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVE
AMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLG
QFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHD
SFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLN
LRDILESDFAFA"
polyA_signal 4490..4545
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4906..4922)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4930..4946)
/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)."
promoter complement(4954..4984)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4999..5020)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5078..5274
/label=SV40 promoter
/note="SV40 early promoter"
polyA_signal 5280..5414
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(5653..6241)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6415..7272)
/label=AmpR
/note="beta-lactamase"
promoter complement(7273..7377)
/label=AmpR promoter
promoter 7405..7421
/label=CAG
/note="CMV early enhancer fused to modified chicken
beta-actin promoter"