pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry vector (V012823)

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V012823 pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry In stock, instant shipping

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Lentiviral vector for Gal4 inducible leucinezipper Pembrolizumab and constitutive mCherry expression

Vector Name:
pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry
Antibiotic Resistance:
Ampicillin
Length:
12269 bp
Type:
Viral Expression & Packaging Vectors
Replication origin:
ori
Source/Author:
Wendell Lim
Promoter:
mPGK
Growth Strain(s):
stbl3
Growth Temperature:
37℃

pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry vector Vector Map

pHR_Gal4UAS_Pembrolizumab_heavychain_T2A_Pembrolizumab_lightchain_PGK_mCherry12269 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000MycT2AHAhIg-kappa-CLPGK promotermCherryWPRE3' LTR (Delta-U3)SP6 promoterpBRrevBampRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRoriL4440SV40 promotersmall t intronSV40 NLSSV40 poly(A) signal5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSLNCX

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V012823                12269 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V012823
VERSION     V012823
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
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             1453..1506
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             1564..1590
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1927..2244
                     /label="hIg-kappa-CL"
                     /note="Human immunoglobulin kappa light chain constant
                     region"
     promoter        2370..2869
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2890..3597
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    3677..4265
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     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(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
                     /label="AmpR promoter"
     CDS             5497..6354
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6528..7116
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     7270..7287
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     promoter        7362..7691
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     intron          8825..8890
                     /label="small t intron"
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9020..9040
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    9465..9599
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             10143..10400
                     /label="5' LTR (truncated)"
                     /note="truncated 5' 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
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             11680..11721
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     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"