pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase vector (V006878)

Price Information

Cat No. Plasmid Name Availability Add to cart
V006878 pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase 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

This is a 3rd generation lentiviral plasmid. It contains both Luciferase and EGFP, with puromycin selection. The promoter is CMV/TO.

Vector Name:
pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase
Antibiotic Resistance:
Ampicillin
Length:
10490 bp
Type:
Mammalian Expression, Lentiviral, Luciferase
Replication origin:
ori
Selection Marker:
Puromycin
Copy Number:
High Copy
Promoter:
hPGK

pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase vector Vector Map

pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase10490 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000CMV enhancerCMV promotertet operatortet operatorattB1EGFPluciferaseattB2V5 taghPGK promoterPuroRWPRE3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7 promoterM13 Forwardf1 oriAmpR promoterAmpRoriL4440CAP binding sitelac promoterlac operatorM13 revT3 promoterRSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTS

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

  • Lorenzatti Hiles G, Cates AL, El-Sawy L, Day KC, Broses LJ, Han AL, Briggs HL, Emamdjomeh A, Chou A, Abel EV, Liebert M, Palmbos PL, Udager AM, Keller ET, Day ML. A surgical orthotopic approach for studying the invasive progression of human bladder cancer. Nat Protoc. 2019 Mar;14(3):738-755.

pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pLentipuro3/TO/V       10490 bp DNA     circular SYN 13-MAY-2021
DEFINITION  3rd generation lentiviral plasmid for expression of EGFP Firefly 
            Luciferase fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10490)
  TITLE     A Surgical Orthotopic Approach to Study the Invasive Progression of 
            Human Bladder Cancer
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10490)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10490)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    510..528
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    531..549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    590..614
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             635..1351
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             1358..3007
                     /label=luciferase
                     /note="firefly luciferase"
     protein_bind    complement(3024..3048)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3101..3142
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        3202..3712
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3734..4330
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    4401..4989
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5069..5302
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5374..5508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5535..5670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5691..5709)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5719..5736)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(5719..5735)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5728..5750)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5877..6332
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6358..6462
                     /label=AmpR promoter
     CDS             6463..7320
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7494..8082
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8236..8253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8370..8391
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8406..8436
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8444..8460
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     8449..8471
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8468..8484
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8468..8484
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8505..8523
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8551..8777
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8778..8958
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9005..9130
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    9623..9856
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10041..10085
                     /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             10234..10275
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    10352..10469
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"