Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012823 | pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
Lentiviral vector for Gal4 inducible leucinezipper Pembrolizumab and constitutive mCherry expression
pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry vector Vector Map
Plasmid Resuspension 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 ℃.
References
- Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH, Walker WJ, McNally KA, Lim WA. Engineering T Cells with Customized Therapeutic Response Programs Using Synthetic Notch Receptors. Cell. 2016 Oct 6;167(2):419-432.e16. doi: 10.1016/j.cell.2016.09.011. Epub 2016 Sep 29.
pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry vector Sequence
LOCUS Exported 12269 bp DNA circular SYN 13-MAY-2021
DEFINITION Lentiviral vector for Gal4 inducible leucinezipper Pembrolizumab and
constitutive mCherry expression.
ACCESSION .
VERSION .
KEYWORDS pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_
Pembrolizumab_lightc...
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12269)
AUTHORS Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH,
Walker WJ, McNally KA, Lim WA
TITLE Engineering T Cells with Customized Therapeutic Response Programs
Using Synthetic Notch Receptors.
JOURNAL Cell. 2016 Oct 6;167(2):419-432.e16. doi:
10.1016/j.cell.2016.09.011. Epub 2016 Sep 29.
PUBMED 27693353
REFERENCE 2 (bases 1 to 12269)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.09.011"; journalName: "Cell"; date: "2016-10-6-
6"; volume: "167"; issue: "2"; pages: "419-432.e16"
FEATURES Location/Qualifiers
source 1..12269
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 58..87
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1453..1506
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 1564..1590
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter 2370..2869
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(2405..2428)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 2802..2821
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 2890..3600
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(3035..3053)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3303..3322
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 3677..4265
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3730..3750)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4148..4159)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4328..4561
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(4586..4604)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4587..4604)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4894..4913)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(5127..5146)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5246..5268
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5306..5324)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5392..5496
/gene="bla"
/label=AmpR promoter
CDS 5497..6357
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5715..5734)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6528..7116
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7017..7036
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7270..7287
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(7357..7377)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7362..7691
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7542..7677
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7604..7623
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(7627..7638)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 8825..8890
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 9020..9040
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 9465..9599
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9502..9521)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9556..9575
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 9767..10400
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 10447..10572
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 11069..11302
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 11487..11531
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 11816..11933
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 12159..12183
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory join(12264..12269,1..4)
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"